Product: PI3K p85/p55 Antibody
Catalog: AF6242
Description: Rabbit polyclonal antibody to PI3K p85/p55
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat, Monkey
Prediction: Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 55kD,85kD; 84kD,54kD(Calculated).
Uniprot: P27986 | Q92569
RRID: AB_2835106

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 100ul $280 In stock
 200ul $350 In stock

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, 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,Monkey
Prediction:
Zebrafish(100%), Bovine(86%), Horse(86%), Sheep(86%), Rabbit(100%), Dog(86%), Chicken(86%), Xenopus(86%)
Clonality:
Polyclonal
Specificity:
PI3K p85/p55 Antibody detects endogenous levels of total PI3K p85/p55.
RRID:
AB_2835106
Cite Format: Affinity Biosciences Cat# AF6242, RRID:AB_2835106.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
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

GRB 1; GRB1; p85 alpha; p85; P85A_HUMAN; Phosphatidylinositol 3 kinase associated p 85 alpha; Phosphatidylinositol 3 kinase regulatory 1; Phosphatidylinositol 3 kinase regulatory subunit alpha; Phosphatidylinositol 3 kinase regulatory subunit polypeptide 1 (p85 alpha); Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha; Phosphatidylinositol 3-kinase regulatory subunit alpha; Phosphoinositide 3 kinase regulatory subunit 1 (alpha); Phosphoinositide 3 kinase regulatory subunit 1 (p85 alpha); Phosphoinositide 3 kinase regulatory subunit 1; Phosphoinositide 3 kinase regulatory subunit polypeptide 1 (p85 alpha); PI3 kinase p85 subunit alpha; PI3-kinase regulatory subunit alpha; PI3-kinase subunit p85-alpha; PI3K; PI3K regulatory subunit alpha; Pik3r1; PtdIns 3 kinase p85 alpha; PtdIns-3-kinase regulatory subunit alpha; PtdIns-3-kinase regulatory subunit p85-alpha; DKFZp686P05226; FLJ41892; OTTHUMP00000009783; OTTHUMP00000009786; p55; p55 gamma; P55G_HUMAN; p55PIK; Phosphatidylinositol 3 kinase regulatory subunit gamma; Phosphatidylinositol 3 kinase regulatory subunit polypeptide 3; Phosphatidylinositol 3 kinase, regulatory subunit, polypeptide 3 (p55, gamma); Phosphatidylinositol 3-kinase 55 kDa regulatory subunit gamma; Phosphatidylinositol 3-kinase regulatory subunit gamma; Phosphoinositide 3 kinase regulatory subunit 3 (gamma); Phosphoinositide 3 kinase regulatory subunit 3; Phosphoinositide 3 kinase regulatory subunit polypeptide 3; Phosphoinositide 3 kinase, regulatory subunit 3 (p55, gamma); Phosphoinositide 3 kinase, regulatory subunit, polypeptide 3 (p55, gamma); PI3 kinase p85 subunit gamma; PI3-kinase regulatory subunit gamma; PI3-kinase subunit p55-gamma; PI3K regulatory subunit gamma; Pik3r3; PtdIns 3 kinase p85 gamma; PtdIns-3-kinase regulatory subunit gamma; PtdIns-3-kinase regulatory subunit p55-gamma;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P27986 P85A_HUMAN:

Isoform 2 is expressed in skeletal muscle and brain, and at lower levels in kidney and cardiac muscle. Isoform 2 and isoform 4 are present in skeletal muscle (at protein level).

Q92569 P55G_HUMAN:

Highest levels in brain and testis. Lower levels in adipose tissue, kidney, heart, lung and skeletal muscle.

Description:
PIK3R1 is a regulatory subunit of phosphoinositide-3-kinase. Mediates binding to a subset of tyrosine-phosphorylated proteins through its SH2 domain. Acts as an adapter, mediating the association of the p110 catalytic unit of the alpha, beta and delta enzymes to the plasma membrane, where p110 phosphorylates inositol lipids. May play an additional role in the regulation of the actin cytoskeleton. Necessary for the insulin-stimulated increase in glucose uptake and glycogen synthesis in insulin-sensitive tissues.
Sequence:
MSAEGYQYRALYDYKKEREEDIDLHLGDILTVNKGSLVALGFSDGQEARPEEIGWLNGYNETTGERGDFPGTYVEYIGRKKISPPTPKPRPPRPLPVAPGSSKTEADVEQQALTLPDLAEQFAPPDIAPPLLIKLVEAIEKKGLECSTLYRTQSSSNLAELRQLLDCDTPSVDLEMIDVHVLADAFKRYLLDLPNPVIPAAVYSEMISLAPEVQSSEEYIQLLKKLIRSPSIPHQYWLTLQYLLKHFFKLSQTSSKNLLNARVLSEIFSPMLFRFSAASSDNTENLIKVIEILISTEWNERQPAPALPPKPPKPTTVANNGMNNNMSLQDAEWYWGDISREEVNEKLRDTADGTFLVRDASTKMHGDYTLTLRKGGNNKLIKIFHRDGKYGFSDPLTFSSVVELINHYRNESLAQYNPKLDVKLLYPVSKYQQDQVVKEDNIEAVGKKLHEYNTQFQEKSREYDRLYEEYTRTSQEIQMKRTAIEAFNETIKIFEEQCQTQERYSKEYIEKFKREGNEKEIQRIMHNYDKLKSRISEIIDSRRRLEEDLKKQAAEYREIDKRMNSIKPDLIQLRKTRDQYLMWLTQKGVRQKKLNEWLGNENTEDQYSLVEDDEDLPHHDEKTWNVGSSNRNKAENLLRGKRDGTFLVRESSKQGCYACSVVVDGEVKHCVINKTATGYGFAEPYNLYSSLKELVLHYQHTSLVQHNDSLNVTLAYPVYAQQRR

MYNTVWSMDRDDADWREVMMPYSTELIFYIEMDPPALPPKPPKPMTSAVPNGMKDSSVSLQDAEWYWGDISREEVNDKLRDMPDGTFLVRDASTKMQGDYTLTLRKGGNNKLIKIYHRDGKYGFSDPLTFNSVVELINHYHHESLAQYNPKLDVKLMYPVSRYQQDQLVKEDNIDAVGKKLQEYHSQYQEKSKEYDRLYEEYTRTSQEIQMKRTAIEAFNETIKIFEEQCHTQEQHSKEYIERFRREGNEKEIERIMMNYDKLKSRLGEIHDSKMRLEQDLKNQALDNREIDKKMNSIKPDLIQLRKIRDQHLVWLNHKGVRQKRLNVWLGIKNEDADENYFINEEDENLPHYDEKTWFVEDINRVQAEDLLYGKPDGAFLIRESSKKGCYACSVVADGEVKHCVIYSTARGYGFAEPYNLYSSLKELVLHYQQTSLVQHNDSLNVRLAYPVHAQMPSLCR

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
Rabbit
100
Zebrafish
100
Horse
86
Bovine
86
Sheep
86
Dog
86
Xenopus
86
Chicken
86
Pig
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P27986/Q92569 As Substrate

Site PTM Type Enzyme
Ubiquitination
S2 Acetylation
Y12 Phosphorylation
S43 Phosphorylation
Y59 Phosphorylation
Y73 Phosphorylation
Y76 Phosphorylation
S83 Phosphorylation P17612 (PRKACA)
T86 Phosphorylation
S147 Phosphorylation
T148 Phosphorylation
T152 Phosphorylation
S154 Phosphorylation
Y203 Phosphorylation P07949 (RET)
S208 Phosphorylation
S265 Phosphorylation
S269 Phosphorylation
S279 Phosphorylation
K346 Ubiquitination
Y368 Phosphorylation P06213 (INSR)
T369 Phosphorylation
K379 Ubiquitination
K419 Ubiquitination
Y426 Phosphorylation
K438 Ubiquitination
K448 Ubiquitination
Y452 Phosphorylation
T454 Phosphorylation
K459 Sumoylation
K459 Ubiquitination
Y463 Phosphorylation
Y467 Phosphorylation
Y470 Phosphorylation
T471 Phosphorylation
T490 Phosphorylation
Y504 Phosphorylation
K506 Ubiquitination
Y508 Phosphorylation P09619 (PDGFRB)
K513 Ubiquitination
R514 Methylation
K519 Ubiquitination
R523 Methylation
Y528 Phosphorylation
K530 Acetylation
S541 Phosphorylation
Y556 Phosphorylation
K567 Ubiquitination
T576 Phosphorylation
Y580 Phosphorylation P06213 (INSR)
T603 Phosphorylation
Y607 Phosphorylation P06213 (INSR) , P12931 (SRC)
S608 Phosphorylation P42336 (PIK3CA) , P68400 (CSNK2A1)
T623 Phosphorylation
S628 Phosphorylation
S629 Phosphorylation
K633 Ubiquitination
S652 Phosphorylation Q15139 (PRKD1)
Y657 Phosphorylation
K674 Ubiquitination
Y679 Phosphorylation
Y688 Phosphorylation
S690 Phosphorylation
Site PTM Type Enzyme
Ubiquitination
K78 Ubiquitination
K106 Acetylation
K111 Ubiquitination
K114 Acetylation
K114 Ubiquitination
K155 Ubiquitination
Y163 Phosphorylation
K170 Ubiquitination
K180 Ubiquitination
Y184 Phosphorylation
S186 Phosphorylation
Y188 Phosphorylation
K191 Ubiquitination
Y195 Phosphorylation
Y199 Phosphorylation
Y202 Phosphorylation
T203 Phosphorylation
T222 Phosphorylation
K224 Ubiquitination
K238 Ubiquitination
Y260 Phosphorylation
S273 Phosphorylation
K274 Ubiquitination
K294 Ubiquitination
K299 Ubiquitination
K319 Ubiquitination
Y341 Phosphorylation P06213 (INSR)
Y373 Phosphorylation
K375 Ubiquitination
K388 Ubiquitination
Y407 Phosphorylation

Research Backgrounds

Function:

Binds to activated (phosphorylated) protein-Tyr kinases, through its SH2 domain, and acts as an adapter, mediating the association of the p110 catalytic unit to the plasma membrane. Necessary for the insulin-stimulated increase in glucose uptake and glycogen synthesis in insulin-sensitive tissues. Plays an important role in signaling in response to FGFR1, FGFR2, FGFR3, FGFR4, KITLG/SCF, KIT, PDGFRA and PDGFRB. Likewise, plays a role in ITGB2 signaling. Modulates the cellular response to ER stress by promoting nuclear translocation of XBP1 isoform 2 in a ER stress- and/or insulin-dependent manner during metabolic overloading in the liver and hence plays a role in glucose tolerance improvement.

PTMs:

Polyubiquitinated in T-cells by CBLB; which does not promote proteasomal degradation but impairs association with CD28 and CD3Z upon T-cell activation.

Phosphorylated. Tyrosine phosphorylated in response to signaling by FGFR1, FGFR2, FGFR3 and FGFR4. Phosphorylated by CSF1R. Phosphorylated by ERBB4. Phosphorylated on tyrosine residues by TEK/TIE2. Dephosphorylated by PTPRJ. Phosphorylated by PIK3CA at Ser-608; phosphorylation is stimulated by insulin and PDGF. The relevance of phosphorylation by PIK3CA is however unclear (By similarity). Phosphorylated in response to KIT and KITLG/SCF. Phosphorylated by FGR.

Tissue Specificity:

Isoform 2 is expressed in skeletal muscle and brain, and at lower levels in kidney and cardiac muscle. Isoform 2 and isoform 4 are present in skeletal muscle (at protein level).

Subunit Structure:

Heterodimer of a regulatory subunit PIK3R1 and a p110 catalytic subunit (PIK3CA, PIK3CB or PIK3CD). Interacts (via SH2 domains) with CCDC88A/GIV (tyrosine-phosphorylated form); the interaction enables recruitment of PIK3R1 to the EGFR receptor, enhancing PI3K activity and cell migration. Interacts (via SH2 domain) with CSF1R (tyrosine phosphorylated). Interacts with PIK3R2; the interaction is dissociated in an insulin-dependent manner (By similarity). Interacts with XBP1 isoform 2; the interaction is direct and induces translocation of XBP1 isoform 2 into the nucleus in a ER stress- and/or insulin-dependent but PI3K-independent manner. Interacts with FER. Interacts (via SH2 domain) with TEK/TIE2 (tyrosine phosphorylated). Interacts with PTK2/FAK1 (By similarity). Interacts with phosphorylated TOM1L1. Interacts with phosphorylated LIME1 upon TCR and/or BCR activation. Interacts with SOCS7. Interacts with RUFY3. Interacts (via SH2 domain) with CSF1R (tyrosine phosphorylated). Interacts with LYN (via SH3 domain); this enhances enzyme activity (By similarity). Interacts with phosphorylated LAT, LAX1 and TRAT1 upon TCR activation. Interacts with CBLB. The SH2 domains interact with the YTHM motif of phosphorylated INSR in vitro. Also interacts with tyrosine-phosphorylated IGF1R in vitro. Interacts with CD28 and CD3Z upon T-cell activation. Interacts with IRS1 and phosphorylated IRS4, as well as with NISCH and HCST. Interacts with FASLG, KIT and BCR. Interacts with AXL, FGFR1, FGFR2, FGFR3 and FGFR4 (phosphorylated). Interacts with FGR and HCK. Interacts with PDGFRA (tyrosine phosphorylated) and PDGFRB (tyrosine phosphorylated). Interacts with ERBB4 (phosphorylated). Interacts with NTRK1 (phosphorylated upon ligand-binding). Interacts with FAM83B; activates the PI3K/AKT signaling cascade. Interacts with APPL1 and APPL2 (By similarity). Interacts with SRC.

(Microbial infection) Interacts with HIV-1 Nef to activate the Nef associated p21-activated kinase (PAK). This interaction depends on the C-terminus of both proteins and leads to increased production of HIV.

(Microbial infection) Interacts with HCV NS5A.

(Microbial infection) Interacts with herpes simplex virus 1 UL46; this interaction activates the PI3K/AKT pathway.

(Microbial infection) Interacts with herpes simplex virus 1 UL46 and varicella virus ORF12; this interaction activates the PI3K/AKT pathway.

Family&Domains:

The SH3 domain mediates the binding to CBLB, and to HIV-1 Nef.

Belongs to the PI3K p85 subunit family.

Function:

Binds to activated (phosphorylated) protein-tyrosine kinases through its SH2 domain and regulates their kinase activity. During insulin stimulation, it also binds to IRS-1.

Tissue Specificity:

Highest levels in brain and testis. Lower levels in adipose tissue, kidney, heart, lung and skeletal muscle.

Subunit Structure:

Heterodimer of a regulatory subunit PIK3R3 and a p110 catalytic subunit (PIK3CA, PIK3CB or PIK3CD). Interacts with AXL.

Family&Domains:

Belongs to the PI3K p85 subunit family.

Research Fields

· 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 > Signaling pathways regulating pluripotency of stem cells.   (View pathway)

· Cellular Processes > Cell motility > Regulation of actin cytoskeleton.   (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 > 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 > Phosphatidylinositol signaling system.

· 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 > AMPK signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Jak-STAT 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 > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Endocrine and metabolic diseases > Type II diabetes mellitus.

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).

· Human Diseases > Infectious diseases: Bacterial > Bacterial invasion of epithelial cells.

· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).

· Human Diseases > Infectious diseases: Parasitic > Amoebiasis.

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

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

· Human Diseases > Infectious diseases: Viral > Measles.

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

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

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

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

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > Proteoglycans 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 > Melanoma.   (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 > Small cell lung cancer.   (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 > Aging > Longevity regulating pathway.   (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway - multiple species.   (View pathway)

· Organismal Systems > Development > Axon guidance.   (View pathway)

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

· Organismal Systems > Immune system > Platelet activation.   (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 > Immune system > Leukocyte transendothelial migration.   (View pathway)

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

· Organismal Systems > Nervous system > Cholinergic synapse.

· Organismal Systems > Sensory system > Inflammatory mediator regulation of TRP channels.   (View pathway)

· 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 > Prolactin signaling pathway.   (View pathway)

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

· Organismal Systems > Endocrine system > Regulation of lipolysis in adipocytes.

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

· Organismal Systems > Excretory system > Aldosterone-regulated sodium reabsorption.

· Organismal Systems > Digestive system > Carbohydrate digestion and absorption.

References

1). ACT001, a novel PAI-1 inhibitor, exerts synergistic effects in combination with cisplatin by inhibiting PI3K/AKT pathway in glioma. Cell Death & Disease, 2019 (PubMed: 31591377) [IF=9.0]

Application: WB    Species: human    Sample: siPAI-1 cells

Fig. 2 |Knockdown of PAI-1 abrogates the migration and invasion ability of glioma cells. i Western blot assay of the control and siPAI-1 cells. Data are represented as the mean ± SEM(error bars) of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001

Application: IHC    Species: mouse    Sample: Tumour

Fig. 7| ACT001 enhances the antitumour effect of cisplatin in vivo. a Xenograft assays in the mice treated with control, ACT001, cisplatin and ACT001 + cisplatin. b Tumour volume changes in the mice treated with control, ACT001, cisplatin and ACT001 + cisplatin. c Body weight changes in the mice treated with control, ACT001, cisplatin and ACT001 + cisplatin. d IHC analysis of the PI3K, p-PI3K, AKT and p-AKT levels. Compared with the control group, the staining of p-PI3K and p-AKT was lighter in the ACT001 treatment groups. In the ACT001 + cisplatin treatment group, the staining of p-PI3K and p-AKT was lighter than in the ACT001-treated group.

2). Loss of PTEN expression is associated with PI3K pathway-dependent metabolic reprogramming in hepatocellular carcinoma. Cell Communication and Signaling, 2020 (PubMed: 32831114) [IF=8.4]

Application: WB    Species: Human    Sample: HHCC cells

Fig. 2 PTEN regulates the development of HCC in vitro and in vivo by inhibiting the activation of PI3K/Akt pathway. HHCC cells were transduced with a lentiviral vector encoding full-length human PTEN (oe-PTEN), with those cells infected with a lentiviral vector harboring empty expression vector as controls (vector-NC). a, Representative Western blots of PI3K, Akt, and mTOR proteins and their quantitation in HHCC cells, normalized to GAPDH. b, EdU-stained cells were captured (× 200) to reflect HHCC cell proliferation. c, Representative Western blots of proliferation markers Ki67 and PCNA and their quantitation in HHCC cells, normalized to GAPDH. d, Wound closure was monitored to measure HHCC cell migration (24 h after scratch). e, HHCC cells invading from Matrigel-coated the upper transwell chamber into the lower one. f, Flow cytometric analysis of PI staining was performed to examine the distribution of HHCC cells throughout G0/G1, S, and G2/M phases of the cell cycle. g, Flow cytometric analysis of Annexin V/PI double staining was performed to determine HHCC cell apoptosis. h, Representative HCC xenograft tumors and the growth of HCC xenograft tumor measured every 7 days in nude mice (n = 6). * p < 0.05 compared with vector-NC by unpaired t-test. Data are shown as mean ± standard deviation of three technical replicates

3). Cathepsin S activity controls chronic stress-induced muscle atrophy and dysfunction in mice. Cellular and Molecular Life Sciences, 2023 (PubMed: 37589754) [IF=8.0]

Application: WB    Species: Mouse    Sample:

Fig. 4 CTSS deficiency ameliorated stress-related anabolic and catabolic molecular alterations. a–e: Representative immunoblotting images and quantitative data for CTSS, IGF-1, IRS-2, p-PI3K, p-Akt, p-mTOR, p-FoxO1α, MuRF-1, MAFbx1, PGC-1α, PPAR-γ, C-caspase-3, and Bcl-2 in GAS muscles at Day 14 after stress (n = 3). Data are mean ± SEM, and p-values were determined by a one-way ANOVA followed by Bonferroni post hoc tests (b–e). CW: CTSS+/+ control mice, CK: CTSS−/− control mice, SW: 14-day-stressed CTSS+/+ mice, SK: 14-day-stressed CTSS−/− mice. *p 

4). Xanthotoxol suppresses non-small cell lung cancer progression and might improve patients' prognosis. PHYTOMEDICINE, 2022 (PubMed: 35932608) [IF=7.9]

5). RPLP2 activates TLR4 in an autocrine manner and promotes HIF-1α-induced metabolic reprogramming in hepatocellular carcinoma. Cell death discovery, 2023 (PubMed: 38052785) [IF=7.0]

Application: WB    Species: Human    Sample: Hep3B and Huh7 cells

Fig. 5 RPLP2 affects HIF-1α by regulating the PI3K/AKT pathway. A KEGG pathway analysis of differentially expressed genes when RPLP2 was knocked down in Hep3B and Huh7 cells. B Western blot analysis showed a decrease in the protein levels of p-PI3K and p-AKT after RPLP2 knockdown and the quantitative analysis (n = 3, mean ± SD) is shown in (C). D After treatment of Hep3B cells with the AKT activator SC79 (10 µM) for 24 h, Western blotting was used to detect the expression of HIF-1α and p-AKT, and quantitative analysis (n = 3, mean ± SD) was performed in (E). After treatment with SC79, the expression of HIF-1α and p-AKT was rescued. F Western blotting was used to determine the effect of SC79 on HIF-1α levels in the cytoplasm and nucleus, and the quantitative analysis (n = 3, mean ± SD) was performed in (G). SC79 restored HIF-1α levels in the cytoplasm and nucleus. H Immunostaining of Hep3B cells with antibodies against HIF-1α (red). Knocking down RPLP2 led to a decrease in the nuclear expression level of HIF-1α, which was abrogated by SC79. Scale bar, 25 μm. *P 

6). β-Caryophyllene promotes the survival of random skin flaps by upregulating the PI3K/AKT signaling pathway. PHYTOMEDICINE, 2024 [IF=6.7]

7). EBV-encoded miRNAs BHRF1-1 and BART2-5p aggravate post- transplant lymphoproliferative disorder via LZTS2-PI3K-AKT axis. Biochemical Pharmacology, 2023 (PubMed: 37419372) [IF=5.8]

8). Metabolomics and integrated network pharmacology analysis reveal attenuates cardiac hypertrophic mechanisms of HuoXin pill. Journal of Ethnopharmacology, 2022 (PubMed: 35304274) [IF=5.4]

9). Tetrahydropalmatine promotes random skin flap survival in rats via the PI3K/AKT signaling pathway. Journal of ethnopharmacology, 2024 (PubMed: 38280663) [IF=5.4]

10). Soluble TREM-1, as a new ligand for the membrane receptor Robo2, promotes hepatic stellate cells activation and liver fibrosis. Journal of Cellular and Molecular Medicine, 2021 (PubMed: 34750987) [IF=5.3]

Application: WB    Species: Human    Sample: LX‐2 cells

FIGURE 4 Liver fibrosis‐related protein expressions under Smad2/3, PI3K/Akt inhibitors in LX‐2 cells (A‐D) Western blot analysis showed the protein expressions of α‐SMA, collagen I, p‐Smad2, p‐Smad3, Samd2/3, p‐PI3K, PI3K, p‐Akt and Akt in control, SIS3/LY294002, sTREM‐1 and sTREM‐1+SIS3/LY294002 groups of LX‐2 cells. ns: not significant, * p < 0.05, ** p < 0.01

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