Phospho-MEK1/2 (Ser218+Ser222/Ser222+Ser226) Antibody - #AF8035

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Product: Phospho-MEK1/2 (Ser218+Ser222/Ser222+Ser226) Antibody
Catalog: AF8035
Description: Rabbit polyclonal antibody to Phospho-MEK1/2 (Ser218+Ser222/Ser222+Ser226)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 45kDa; 43kD,44kD(Calculated).
Uniprot: Q02750 | P36507
RRID: AB_2840098

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 100ul $350 In stock
 200ul $450 In stock

Lead Time: Same day delivery

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:1000-3000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-MEK1/2 (Ser218+Ser222/Ser222+Ser226) Antibody detects endogenous levels of MEK1/2 only when phosphorylated at S218+S222(MEK1)/S222+S226(MEK2).
RRID:
AB_2840098
Cite Format: Affinity Biosciences Cat# AF8035, RRID:AB_2840098.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Dual specificity mitogen activated protein kinase kinase 1; Dual specificity mitogen-activated protein kinase kinase 1; ERK activator kinase 1; MAP kinase kinase 1; MAP kinase/Erk kinase 1; MAP2K1; MAPK/ERK kinase 1; MAPKK 1; MAPKK1; MEK 1; Mek1; MEKK1; Mitogen activated protein kinase kinase 1; MKK 1; MKK1; MP2K1_HUMAN; PRKMK1; Protein kinase mitogen activated kinase 1 (MAP kinase kinase 1); Protein kinase mitogen activated, kinase 1; Cardiofaciocutaneous syndrome; CFC syndrome; CFC4; Dual specificity mitogen activated protein kinase kinase 2; Dual specificity mitogen-activated protein kinase kinase 2; ERK activator kinase 2; FLJ26075; MAP kinase kinase 2; map2k2; MAPK / ERK kinase 2; MAPK/ERK kinase 2; MAPKK 2; MAPKK2; MEK 2; MEK2; Microtubule associated protein kinase kinase 2; Mitogen activated protein kinase kinase 2; Mitogen activated protein kinase kinase 2 p45; MKK 2; MKK2; MP2K2_HUMAN; OTTHUMP00000165826; OTTHUMP00000165827; PRKMK 2; PRKMK2;

Immunogens

Immunogen:
Uniprot:

Q02750(MP2K1_HUMAN) >>Visit HPA database.

P36507(MP2K2_HUMAN) >>Visit HPA database.

Gene(ID):
MAP2K1(5604) | MAP2K2(5605)
Expression:
Q02750 MP2K1_HUMAN:

Widely expressed, with extremely low levels in brain.

Sequence:
MPKKKPTPIQLNPAPDGSAVNGTSSAETNLEALQKKLEELELDEQQRKRLEAFLTQKQKVGELKDDDFEKISELGAGNGGVVFKVSHKPSGLVMARKLIHLEIKPAIRNQIIRELQVLHECNSPYIVGFYGAFYSDGEISICMEHMDGGSLDQVLKKAGRIPEQILGKVSIAVIKGLTYLREKHKIMHRDVKPSNILVNSRGEIKLCDFGVSGQLIDSMANSFVGTRSYMSPERLQGTHYSVQSDIWSMGLSLVEMAVGRYPIPPPDAKELELMFGCQVEGDAAETPPRPRTPGRPLSSYGMDSRPPMAIFELLDYIVNEPPPKLPSGVFSLEFQDFVNKCLIKNPAERADLKQLMVHAFIKRSDAEEVDFAGWLCSTIGLNQPSTPTHAAGV

MLARRKPVLPALTINPTIAEGPSPTSEGASEANLVDLQKKLEELELDEQQKKRLEAFLTQKAKVGELKDDDFERISELGAGNGGVVTKVQHRPSGLIMARKLIHLEIKPAIRNQIIRELQVLHECNSPYIVGFYGAFYSDGEISICMEHMDGGSLDQVLKEAKRIPEEILGKVSIAVLRGLAYLREKHQIMHRDVKPSNILVNSRGEIKLCDFGVSGQLIDSMANSFVGTRSYMAPERLQGTHYSVQSDIWSMGLSLVELAVGRYPIPPPDAKELEAIFGRPVVDGEEGEPHSISPRPRPPGRPVSGHGMDSRPAMAIFELLDYIVNEPPPKLPNGVFTPDFQEFVNKCLIKNPAERADLKMLTNHTFIKRSEVEEVDFAGWLCKTLRLNQPGTPTRTAV

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Xenopus
100
Zebrafish
100
Chicken
100
Rabbit
100
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q02750/P36507 As Substrate

Site PTM Type Enzyme
M1 Acetylation
K6 Ubiquitination
T13 Acetylation
S23 Phosphorylation
T25 Phosphorylation
S26 Phosphorylation P36507 (MAP2K2)
S30 Phosphorylation
K40 Ubiquitination
K51 Ubiquitination
T59 Phosphorylation O14757 (CHEK1)
K61 Ubiquitination
K63 Ubiquitination
K68 Ubiquitination
K88 Ubiquitination
S94 Phosphorylation
K101 Ubiquitination
K108 Sumoylation
K108 Ubiquitination
K163 Ubiquitination
K196 Ubiquitination
S198 Phosphorylation
K209 Ubiquitination
S216 Phosphorylation
S222 Acetylation
S222 Phosphorylation P15056 (BRAF) , P41279 (MAP3K8) , O15530 (PDPK1) , P53350 (PLK1)
S226 Acetylation
S226 Phosphorylation O15530 (PDPK1) , P53350 (PLK1) , P15056 (BRAF) , P41279 (MAP3K8) , P36507 (MAP2K2)
T230 Phosphorylation
S293 Phosphorylation
S295 Phosphorylation
S306 Phosphorylation
S312 Phosphorylation
K352 Ubiquitination
K361 Ubiquitination
K370 Ubiquitination
S372 Phosphorylation
K385 Ubiquitination
T394 Phosphorylation
T396 Phosphorylation P36507 (MAP2K2)
T398 Phosphorylation
Site PTM Type Enzyme
T23 Phosphorylation
S24 Phosphorylation Q6VAB6 (KSR2)
S25 Phosphorylation
K36 Ubiquitination
T55 Phosphorylation O14757 (CHEK1)
K57 Ubiquitination
K64 Ubiquitination
K70 Ubiquitination
S72 Phosphorylation Q6VAB6 (KSR2)
K88 Ubiquitination
K97 Ubiquitination
K104 Sumoylation
K104 Ubiquitination
K168 Ubiquitination
K175 Acetylation
K175 Ubiquitination
K192 Ubiquitination
S194 Phosphorylation
K205 Ubiquitination
S212 Phosphorylation
S218 Phosphorylation P53350 (PLK1) , O15530 (PDPK1) , P15056 (BRAF) , P41279 (MAP3K8) , Q5TCX8 (MAP3K21) , P00540 (MOS) , Q13233 (MAP3K1) , P10398 (ARAF) , P04049 (RAF1)
S222 Phosphorylation Q5TCX8 (MAP3K21) , P10398 (ARAF) , Q13233 (MAP3K1) , P53350 (PLK1) , O15530 (PDPK1) , P00540 (MOS) , P04049 (RAF1) , P15056 (BRAF) , P41279 (MAP3K8)
T226 Phosphorylation
S231 Phosphorylation
S244 Phosphorylation P41279 (MAP3K8)
S248 Phosphorylation P41279 (MAP3K8)
T286 Phosphorylation Q00535 (CDK5) , P06493 (CDK1) , P27361 (MAPK3) , P28482 (MAPK1)
T292 Phosphorylation P06493 (CDK1) , P27361 (MAPK3) , Q00535 (CDK5) , P28482 (MAPK1)
S298 Phosphorylation Q13153 (PAK1) , Q02750 (MAP2K1) , O75914 (PAK3)
S299 Phosphorylation
Y300 Phosphorylation Q02750 (MAP2K1)
K344 Ubiquitination
K353 Ubiquitination
K362 Acetylation
S385 Phosphorylation
T386 Phosphorylation P28482 (MAPK1) , P27361 (MAPK3)
T388 Phosphorylation

PTMs - Q02750/P36507 As Enzyme

Substrate Site Source
P05787 (KRT8) S74 Uniprot
P27361 (MAPK3) T202 Uniprot
P27361 (MAPK3) Y204 Uniprot
P28482 (MAPK1) T185 Uniprot
P28482 (MAPK1) Y187 Uniprot
P36507 (MAP2K2) S26 Uniprot
P36507 (MAP2K2) S226 Uniprot
P36507 (MAP2K2) T396 Uniprot
P55211-1 (CASP9) T125 Uniprot
Substrate Site Source
O75676-1 (RPS6KA4) S196 Uniprot
O75676-1 (RPS6KA4) S343 Uniprot
O75676-1 (RPS6KA4) S347 Uniprot
O75676-1 (RPS6KA4) S360 Uniprot
O75676-2 (RPS6KA4) T562 Uniprot
O75676-1 (RPS6KA4) T568 Uniprot
P04049 (RAF1) S338 Uniprot
P05787 (KRT8) S74 Uniprot
P15172 (MYOD1) Y156 Uniprot
P17542 (TAL1) S122 Uniprot
P27361 (MAPK3) T202 Uniprot
P27361 (MAPK3) Y204 Uniprot
P27361 (MAPK3) T207 Uniprot
P27361 (MAPK3) Y210 Uniprot
P28482 (MAPK1) T185 Uniprot
P28482 (MAPK1) Y187 Uniprot
P32121 (ARRB2) T382 Uniprot
P35568 (IRS1) S307 Uniprot
P37231 (PPARG) S112 Uniprot
P49841 (GSK3B) Y216 Uniprot
P55211-1 (CASP9) T125 Uniprot
Q00613 (HSF1) S326 Uniprot
Q02750 (MAP2K1) S298 Uniprot
Q02750 (MAP2K1) Y300 Uniprot
Q02750-1 (MAP2K1) S304 Uniprot

Research Backgrounds

Function:

Dual specificity protein kinase which acts as an essential component of the MAP kinase signal transduction pathway. Binding of extracellular ligands such as growth factors, cytokines and hormones to their cell-surface receptors activates RAS and this initiates RAF1 activation. RAF1 then further activates the dual-specificity protein kinases MAP2K1/MEK1 and MAP2K2/MEK2. Both MAP2K1/MEK1 and MAP2K2/MEK2 function specifically in the MAPK/ERK cascade, and catalyze the concomitant phosphorylation of a threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located in the extracellular signal-regulated kinases MAPK3/ERK1 and MAPK1/ERK2, leading to their activation and further transduction of the signal within the MAPK/ERK cascade. Activates BRAF in a KSR1 or KSR2-dependent manner; by binding to KSR1 or KSR2 releases the inhibitory intramolecular interaction between KSR1 or KSR2 protein kinase and N-terminal domains which promotes KSR1 or KSR2-BRAF dimerization and BRAF activation. Depending on the cellular context, this pathway mediates diverse biological functions such as cell growth, adhesion, survival and differentiation, predominantly through the regulation of transcription, metabolism and cytoskeletal rearrangements. One target of the MAPK/ERK cascade is peroxisome proliferator-activated receptor gamma (PPARG), a nuclear receptor that promotes differentiation and apoptosis. MAP2K1/MEK1 has been shown to export PPARG from the nucleus. The MAPK/ERK cascade is also involved in the regulation of endosomal dynamics, including lysosome processing and endosome cycling through the perinuclear recycling compartment (PNRC), as well as in the fragmentation of the Golgi apparatus during mitosis.

PTMs:

Phosphorylation at Ser-218 and Ser-222 by MAP kinase kinase kinases (BRAF or MEKK1) positively regulates kinase activity. Also phosphorylated at Thr-292 by MAPK1/ERK2 and at Ser-298 by PAK. MAPK1/ERK2 phosphorylation of Thr-292 occurs in response to cellular adhesion and leads to inhibition of Ser-298 phosphorylation by PAK.

Acetylation by Yersinia yopJ prevents phosphorylation and activation, thus blocking the MAPK signaling pathway.

Subcellular Location:

Cytoplasm>Cytoskeleton>Microtubule organizing center>Centrosome. Cytoplasm>Cytoskeleton>Microtubule organizing center>Spindle pole body. Cytoplasm. Nucleus. Membrane>Peripheral membrane protein.
Note: Localizes at centrosomes during prometaphase, midzone during anaphase and midbody during telophase/cytokinesis (PubMed:14737111). Membrane localization is probably regulated by its interaction with KSR1 (PubMed:10409742).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Widely expressed, with extremely low levels in brain.

Subunit Structure:

Found in a complex with at least BRAF, HRAS, MAP2K1, MAPK3/ERK1 and RGS14 (By similarity). Forms a heterodimer with MAP2K2/MEK2 (By similarity). Forms heterodimers with KSR2 which further dimerize to form tetramers (By similarity). Interacts with KSR1 or KSR2 and BRAF; the interaction with KSR1 or KSR2 mediates KSR1-BRAF or KSR2-BRAF dimerization. Interacts with ARBB2, LAMTOR3, MAPK1/ERK2 and RAF1 (By similarity). Interacts with MORG1 (By similarity). Interacts with PPARG. Interacts with isoform 1 of VRK2. Interacts with SGK1. Interacts with BIRC6/bruce. Interacts with KAT7; the interaction promotes KAT7 phosphorylation (By similarity).

(Microbial infection) Interacts with Yersinia yopJ.

Family&Domains:

The proline-rich region localized between residues 270 and 307 is important for binding to RAF1 and activation of MAP2K1/MEK1.

Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase subfamily.

Function:

Catalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located in MAP kinases. Activates the ERK1 and ERK2 MAP kinases (By similarity). Activates BRAF in a KSR1 or KSR2-dependent manner; by binding to KSR1 or KSR2 releases the inhibitory intramolecular interaction between KSR1 or KSR2 protein kinase and N-terminal domains which promotes KSR1 or KSR2-BRAF dimerization and BRAF activation.

PTMs:

MAPKK is itself dependent on Ser/Thr phosphorylation for activity catalyzed by MAP kinase kinase kinases (RAF or MEKK1). Phosphorylated by MAP2K1/MEK1 (By similarity).

Acetylation of Ser-222 and Ser-226 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the MAPK signaling pathway.

Subcellular Location:

Cytoplasm. Membrane>Peripheral membrane protein.
Note: Membrane localization is probably regulated by its interaction with KSR1.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Subunit Structure:

Interacts with MORG1 (By similarity). Interacts with SGK1. Interacts with KSR1. Interacts with KSR1 and BRAF; the interaction with KSR1 mediates KSR1-BRAF dimerization. Interacts with GLS.

Family&Domains:

Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase subfamily.

Research Fields

· Cellular Processes > Cell growth and death > Oocyte meiosis.   (View pathway)

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

· Cellular Processes > Cell growth and death > Apoptosis.   (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Gap junction.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Signaling pathways regulating pluripotency of stem cells.   (View pathway)

· Cellular Processes > Cell motility > Regulation of actin cytoskeleton.   (View pathway)

· Environmental Information Processing > Signal transduction > MAPK signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > ErbB signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Ras signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Rap1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > cAMP signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > FoxO signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Phospholipase D signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > mTOR signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Apelin signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > TNF signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Neurodegenerative diseases > Prion diseases.

· Human Diseases > Substance dependence > Alcoholism.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Influenza A.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Colorectal cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Renal cell carcinoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Pancreatic cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Endometrial cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Glioma.   (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Thyroid cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Melanoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Bladder cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia.   (View pathway)

· Human Diseases > Cancers: Specific types > Acute myeloid leukemia.   (View pathway)

· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Breast cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Central carbon metabolism in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Choline metabolism in cancer.   (View pathway)

· Organismal Systems > Immune system > Chemokine signaling pathway.   (View pathway)

· Organismal Systems > Circulatory system > Vascular smooth muscle contraction.   (View pathway)

· Organismal Systems > Development > Osteoclast differentiation.   (View pathway)

· Organismal Systems > Immune system > Toll-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Natural killer cell mediated cytotoxicity.   (View pathway)

· Organismal Systems > Immune system > T cell receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > B cell receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Fc epsilon RI signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Fc gamma R-mediated phagocytosis.   (View pathway)

· Organismal Systems > Nervous system > Long-term potentiation.

· Organismal Systems > Nervous system > Neurotrophin signaling pathway.   (View pathway)

· Organismal Systems > Nervous system > Cholinergic synapse.

· Organismal Systems > Nervous system > Serotonergic synapse.

· Organismal Systems > Nervous system > Long-term depression.

· Organismal Systems > Endocrine system > Insulin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Progesterone-mediated oocyte maturation.

· Organismal Systems > Endocrine system > Estrogen signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Melanogenesis.

· Organismal Systems > Endocrine system > Prolactin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

References

1). LncRNA CD27-AS1 promotes acute myeloid leukemia progression through the miR-224-5p/PBX3 signaling circuit. Cell Death & Disease, 2021 (PubMed: 34006845) [IF=9.0]

Application: WB    Species: Human    Sample: AML cells

Fig. 5 CD27-AS1 regulates MAPK signaling pathway in the AML cell lines. a HL-60 and KG-1 cells were infected with LV-CD27-AS1 and LV-CD27- AS1-Sh1. Protein levels of P38, p-P38, JNK, p-JNK, p-C-raf, p-MEK1/2, ERK, and p-ERK were detected using western blotting. b Densitometry analysis of protein levels in both cells was performed. c U0126, a MEK1/2 inhibitor, was used to treat the AML cells with CD27-AS1 upregulation. Relative protein levels of p-ERK and ERK were measured using western blotting. d CCK-8 assay was performed to check cell viability. N = 3. Data were shown as means ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001.
2). Serum proteomics analysis based on label-free revealed the protective effect of Chinese herbal formula Gu-Ben-Fang-Xiao. BIOMEDICINE & PHARMACOTHERAPY, 2019 (PubMed: 31520916) [IF=7.5]

Application: WB    Species: mouse    Sample: lung

Figure S2. |The effect of GBFXD on RAF/MEK/ERK signaling pathway. Western blotting was used to detect the relative protein expressions of P-RAF/RAF, P-MEK/MEK, and P-ERK/ERK.
3). Intra-Pancreatic Insulin Nourishes Cancer Cells: Do Insulin-Receptor Antagonists such as PGG and EGCG Play a Role?. The American Journal of Chinese Medicine, 2020 (PubMed: 32468825) [IF=5.7]

Application: WB    Species: Mice    Sample: tumors tissue

Figure 4. See the legend of Fig. 3 for study design. (A, B) t-Akt and p-Akt results from the sub-studies that involved PGG (A) and EGCG (B). (C, D) t-MEK and p-MEK results from the sub-studies that involved PGG (C) and EGCG (D). (E, F) t-ERK1/2 and p-ERK1/2 results from the sub-studies that involved PGG (E) and EGCG (F). β-Actin was used as a loading control. The blotting data are derived from representative tumors. The histograms include data from all tumors. *p < 0:05.
4). Wenshen-Jianpi prescription, a Chinese herbal medicine, improves visceral hypersensitivity in a rat model of IBS-D by regulating the MEK/ERK signal pathway. Frontiers in Pharmacology, 2022 (PubMed: 36210803) [IF=5.6]

Application: WB    Species: Rat    Sample: colon tissues

FIGURE 7 Effect of WJP on the relative expression of mRNA of MEK1, MEK2, ERK1, and ERK2 and the relative expression of protein of p-MEK1/2, p-ERK1, and p-ERK2 in the colon in the four groups. (A) The relative expression of mRNA of MEK1, MEK2, ERK1, and ERK2. (B) The relative expression of protein of p-MEK1/2. (C) The relative expression of protein of p-ERK1 and p-ERK2. (D) Western blot analysis. *p < 0.05; **p < 0.01, versus the control group. # p < 0.05; ## p < 0.01, versus the model group. ▲ p < 0.05; ▲▲ p < 0.01, versus the positive drug group.
5). l-Theanine attenuates neointimal hyperplasia via suppression of vascular smooth muscle cell phenotypic modulation. JOURNAL OF NUTRITIONAL BIOCHEMISTRY, 2020 (PubMed: 32402912) [IF=5.6]

Application: WB    Species: Rat    Sample: VSMCs

Fig. 5. L-Theanine inhibited activation of ERK1/2 but not STAT3 induced by PDGF-BB and AngII in VSMCs. (a, c) VSMCs were stimulated with PDGF-BB or AngII for the indicated times with or without L-theanine (200 μM). The protein levels of phospho-ERK1/2, ERK, phospho-STAT3, STAT3 were determined using western blot analysis. For (a), *P(5 min)=0.034, *P(10 min)=0.028, **P(30 min)=0.002, compared to the control group; for (c), *P(5 min)=0.041, **P(10 min)=0.008, **P(15 min)=0.008, **P(30 min)=0.016, compared with the control group. ***Pb.001, compared with control group; ns, no significance. (b, d) VSMCs were pretreated with L-theanine (0, 100, 200, 400 μM) for 1 h and then stimulated with PDGF-BB (20 ng/ ml) or AngII (100 nM) for 10 min. The protein levels of phospho-ERK1/2, ERK, phospho-STAT3, STAT3 were determined by using western blot analysis. For (b), **P(L-The 100 μM)= 0.004, compared with the PDGF-BB group; for (d), *P(L-The 100 μM)=0.024, **P(L-The 200 μM)=0.003, compared with the AngII group. ###Pb.001 compared with control group; ***Pb.001, compared with PDGF-BB or AngII group; ns, no significance. Data of 3 independent experiments performed in duplicates are presented as mean±S.E.M. Data were analyzed by two-way ANOVA (a, c) and one-way ANOVA (b, d) with Fisher's LSD test.
6). β-Patchoulene Ameliorates Water Transport and the Mucus Barrier in 5-Fluorouracil-Induced Intestinal Mucositis Rats via the cAMP/PKA/CREB Signaling Pathway. Frontiers in Pharmacology, 2021 (PubMed: 34512326) [IF=5.6]

Application: WB    Species: Rat    Sample:

FIGURE 5 Effects of β-PAE on AQP3 expression and cAMP/PKA/CREB signaling pathway-related proteins. (A,B) AQP3 expression; (C,D) VIP, VIPR2, cAMP and PKA expression; (E,F) MEK1/2, p-MEK1/2, ERK, p-ERK, p-p38, p38, MSK1, p-MSK1, CREB, p-CREB, and P300/CBP expression. Data are shown as mean ± SD (n = 3). # p < 0.05, ## p < 0.01 versus control group; * p < 0.05, ** p < 0.01 versus 5-FU group.
7). Loganin alleviates sepsis-induced acute lung injury by regulating macrophage polarization and inhibiting NLRP3 inflammasome activation. International Immunopharmacology, 2021 (PubMed: 33744777) [IF=5.6]

Application: WB    Species: Mice    Sample: lung tissues

Fig. 5. Loganin inhibits ERK and NF-κB signaling in sepsis-induced ALI mice. (A) The levels of pro-inflammatory cytokines TNF-α and IL-6 in BALF were measured by ELISA assay. (B) The expression of Bcl-2 and Bcl-xl were detected by western blotting. (C) The expression of MEK, p-MEK (Ser217/ Ser221), ERK, p-ERK (Thr202/Tyr204), IKK-α/β, p-IKK-α/β (Ser180/Ser181), IκB-α, p-IκB-α (Ser32/36), NF-κB p65 (cytoplasm), NF-κB p65 (nucleus) were determined by western blotting. Data were shown as means ± SD. n = 6. *p < 0.05 vs. sham. #p < 0.05 vs. CLP. ERK, extra-cellular signal-regulated kinase; NF-κB, nuclear factor-kappa B; TNF-α, tumor necrosis factor-α; IL-6, interleukin-6; BALF, bronchoalveolar lavage fluid; CLP, cecal ligation and puncture.
8). Circ_0011385 knockdown inhibits cell proliferation, migration and invasion, whereas promotes cell apoptosis by regulating miR-330-3p/MYO6 axis in colorectal cancer. Biomedical Journal, 2023 (PubMed: 35091088) [IF=5.5]

Application: WB    Species: Human    Sample: HCT116 and SW480 cells

Fig. 4. Circ_0011385 silencing suppressed CRC development by interacting with miR-330-3p. (A and B) The effect of miR-330-3p inhibitor on miR-330-3p expression was determined by qRT-PCR in HCT116 and SW480 cells. (C–E) The impacts between circ_0011385 silencing and miR-330-3p inhibitor on the proliferation of HCT116 and SW480 cells were unveiled by MTT assay and cell colony formation assay. (F, I and J) Flow cytometry analysis was utilized to illustrate the effects between circ_0011385 silencing and miR-330-3p inhibitor on the apoptosis and cell cycle of HCT116 and SW480 cells. (G and H) Wound-healing and transwell assays were performed to reveal the influences between circ_0011385 absence and miR-330-3p inhibitor on the migration and invasion of HCT116 and SW480 cells, respectively. (K and L) Western blot assay was conducted to determine the influences between circ_0011385 downregulation and miR-330-3p inhibitor on the protein expression of CDK1 and Cyclin D1 in HCT116 and SW480 cells.
9). Optimized new Shengmai powder ameliorates myocardial fibrosis in rats with heart failure by inhibition of the MAPK signaling pathway. Journal of ethnopharmacology, 2024 (PubMed: 37739104) [IF=5.4]

Application: WB    Species: Rat    Sample:

Fig. 7. Effects of ONSMP on ERK1/2 signaling pathway. (A) Representative western blotting bands of p-RAS, RAS, p-RAF, RAF, p-MEK1/2, MEK1/2, p-ERK1/2, ERK1/2, p-ELK, ELK, and c-Fos in the myocardium, with vinculin as internal reference. (B–G) Statistical graphs of p-RAS/RAS, p-RAF/RAF, p-MEK1/2/MEK1/2, p-ERK1/2/ERK1/2, p-ELK/ELK, and c-Fos (n = 3). Data are presented as mean ± SEM. nsP > 0.05; ▲▲P < 0.01 vs the sham; **P < 0.01 vs the model; #P < 0.05, ##P < 0.01 vs the ONSMP-L; ※P < 0.05, ※※P < 0.01 vs the ONSMP-M.
10). Mechanisms of pancreatic tumor suppression mediated by Xiang-lian pill: An integrated in silico exploration and experimental validation. JOURNAL OF ETHNOPHARMACOLOGY, 2022 (PubMed: 35931303) [IF=5.4]

Application: WB    Species: Mouse    Sample:

Fig. 6. XLP suppressed pancreatic tumor growth of tumor-bearing mice. (A) Tumors of mice in different groups treated with different doses of XLP; (B) Tumor weights among different groups; (C) Tumor volume increased over time; (D) HE staining of mouse liver and kidney tissues; (E) Mouse body weight over time; (F) Tumor pathology among four groups under microscopy (HE, 200 × ), and the expression of Ki-67, PTGS2 and PTGS1 among four groups (IHC, 400 × ); (G) Quantitation of the IHC positive area of Ki-67, PTGS2 and PTGS1 in the tumor tissues of four groups (n = 8 each group, each dot represents one sample); (H) The expression of PTGS1 and PTGS2, the phosphorylation of MEK and ERK among four groups by immunoblotting; (I) Statistical analysis of the immunoblotting results. * denotes p < 0.05, ** p < 0.01 and *** p < 0.001 when compared with the control group; # denotes p < 0.05, ## p < 0.01 and ### p < 0.001 when compared with the 0.78 g/kg XLP group.
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