Product: IKK-beta Antibody
Catalog: AF6009
Source: Rabbit
Application: WB, IHC, IF/ICC, ELISA(peptide)
Reactivity: Human, Mouse, Rat, Monkey
Prediction: Pig, Zebrafish, Bovine, Horse, Rabbit, Dog, Xenopus
Mol.Wt.: 86kD; 87kD(Calculated).
Uniprot: O14920
RRID: AB_2834943

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 100ul $280 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, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.
Reactivity:
Human,Mouse,Rat,Monkey
Prediction:
Pig(100%), Zebrafish(100%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
IKK-beta Antibody detects endogenous levels of total IKK-beta.
RRID:
AB_2834943
Cite Format: Affinity Biosciences Cat# AF6009, RRID:AB_2834943.
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

I kappa B kinase 2; I kappa B kinase beta; I-kappa-B kinase 2; I-kappa-B-kinase beta; IkBKB; IKK beta; IKK-B; IKK-beta; IKK2; IKKB; IKKB_HUMAN; IMD15; Inhibitor of kappa light polypeptide gene enhancer in B cells, kinase beta; Inhibitor of nuclear factor kappa-B kinase subunit beta; NFKBIKB; Nuclear factor NF-kappa-B inhibitor kinase beta;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
O14920 IKKB_HUMAN:

Highly expressed in heart, placenta, skeletal muscle, kidney, pancreas, spleen, thymus, prostate, testis and peripheral blood.

Description:
IKK-beta is a kinase of the IKK family. Phosphorylates inhibitors of NF-kappa-B thus leading to the dissociation of the inhibitor/NF-kappa-B complex and ultimately the degradation of the inhibitor. Preferentially found as a heterodimer with IKK-alpha but also as an homodimer.
Sequence:
MSWSPSLTTQTCGAWEMKERLGTGGFGNVIRWHNQETGEQIAIKQCRQELSPRNRERWCLEIQIMRRLTHPNVVAARDVPEGMQNLAPNDLPLLAMEYCQGGDLRKYLNQFENCCGLREGAILTLLSDIASALRYLHENRIIHRDLKPENIVLQQGEQRLIHKIIDLGYAKELDQGSLCTSFVGTLQYLAPELLEQQKYTVTVDYWSFGTLAFECITGFRPFLPNWQPVQWHSKVRQKSEVDIVVSEDLNGTVKFSSSLPYPNNLNSVLAERLEKWLQLMLMWHPRQRGTDPTYGPNGCFKALDDILNLKLVHILNMVTGTIHTYPVTEDESLQSLKARIQQDTGIPEEDQELLQEAGLALIPDKPATQCISDGKLNEGHTLDMDLVFLFDNSKITYETQISPRPQPESVSCILQEPKRNLAFFQLRKVWGQVWHSIQTLKEDCNRLQQGQRAAMMNLLRNNSCLSKMKNSMASMSQQLKAKLDFFKTSIQIDLEKYSEQTEFGITSDKLLLAWREMEQAVELCGRENEVKLLVERMMALQTDIVDLQRSPMGRKQGGTLDDLEEQARELYRRLREKPRDQRTEGDSQEMVRLLLQAIQSFEKKVRVIYTQLSKTVVCKQKALELLPKVEEVVSLMNEDEKTVVRLQEKRQKELWNLLKIACSKVRGPVSGSPDSMNASRLSQPGQLMSQPSTASNSLPEPAKKSEELVAEAHNLCTLLENAIQDTVREQDQSFTALDWSWLQTEEEEHSCLEQAS

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
Dog
100
Xenopus
100
Zebrafish
100
Rabbit
100
Sheep
69
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O14920 As Substrate

Site PTM Type Enzyme
S4 Phosphorylation
S6 Phosphorylation
T23 Phosphorylation
K44 Ubiquitination
K106 Ubiquitination
K147 Ubiquitination
K163 Ubiquitination
Y169 Phosphorylation
K171 Ubiquitination
S177 Phosphorylation P54646 (PRKAA2) , PR:000035736 (hMAP3K7/Phos:1) , P17252 (PRKCA) , Q02156 (PRKCE) , O43318 (MAP3K7) , Q99558 (MAP3K14) , Q05513 (PRKCZ) , Q9UHD2 (TBK1)
C179 S-Nitrosylation
T180 Acetylation
T180 Phosphorylation
S181 Phosphorylation Q9UHD2 (TBK1) , P17252 (PRKCA) , O00141 (SGK1) , O15530 (PDPK1) , Q99558 (MAP3K14) , P54646 (PRKAA2) , Q9Y6K9 (IKBKG) , PR:000035736 (hMAP3K7/Phos:1) , O14920 (IKBKB) , O43318 (MAP3K7) , Q05513 (PRKCZ)
Y188 Phosphorylation P12931 (SRC) , P07948 (LYN)
Y199 Phosphorylation P12931 (SRC) , P07948 (LYN)
S239 Phosphorylation
S246 Phosphorylation
S256 Phosphorylation
S257 Phosphorylation
S258 Phosphorylation
Y261 Phosphorylation
S267 Phosphorylation
K301 Ubiquitination
T324 Phosphorylation
Y325 Phosphorylation
S332 Phosphorylation
S335 Phosphorylation
K337 Ubiquitination
T344 Phosphorylation
K365 Ubiquitination
T368 Phosphorylation
S393 Phosphorylation
T396 Phosphorylation
Y397 Phosphorylation
T399 Phosphorylation
S402 Phosphorylation
S409 Phosphorylation
S411 Phosphorylation
K418 Ubiquitination
K467 Ubiquitination
S471 Phosphorylation
S474 Phosphorylation
S476 Phosphorylation
T488 Phosphorylation
S489 Phosphorylation
S507 Phosphorylation
K509 Ubiquitination
S550 Phosphorylation
K555 Ubiquitination
T559 Phosphorylation
T583 Phosphorylation
S587 Phosphorylation
S600 Phosphorylation
K603 Ubiquitination
Y609 Phosphorylation
T610 Phosphorylation
K628 Ubiquitination
S634 Phosphorylation
K652 Ubiquitination
K659 Acetylation
K664 Ubiquitination
S670 Phosphorylation O14920 (IKBKB)
S672 Phosphorylation O14920 (IKBKB)
S675 Phosphorylation O14920 (IKBKB)
S679 Phosphorylation O14920 (IKBKB)
S682 Phosphorylation O14920 (IKBKB)
S689 Phosphorylation O14920 (IKBKB)
S692 Phosphorylation O14920 (IKBKB)
T693 Phosphorylation
S695 Phosphorylation O14920 (IKBKB)
S697 Phosphorylation O14920 (IKBKB)
K703 Ubiquitination
S705 Phosphorylation O14920 (IKBKB)
S733 O-Glycosylation
S733 Phosphorylation O14920 (IKBKB) , P53350 (PLK1)
T735 Phosphorylation
S740 Phosphorylation O14920 (IKBKB) , P53350 (PLK1)
S750 Phosphorylation O14920 (IKBKB) , P53350 (PLK1)
S756 Phosphorylation O14920 (IKBKB)

PTMs - O14920 As Enzyme

Substrate Site Source
O14920-1 (IKBKB) C179 Uniprot
O14920 (IKBKB) S181 Uniprot
O14920 (IKBKB) S670 Uniprot
O14920 (IKBKB) S672 Uniprot
O14920 (IKBKB) S675 Uniprot
O14920-1 (IKBKB) S679 Uniprot
O14920 (IKBKB) S682 Uniprot
O14920 (IKBKB) S689 Uniprot
O14920-1 (IKBKB) S692 Uniprot
O14920 (IKBKB) S695 Uniprot
O14920-1 (IKBKB) S697 Uniprot
O14920-1 (IKBKB) S705 Uniprot
O14920-1 (IKBKB) S733 Uniprot
O14920 (IKBKB) S740 Uniprot
O14920-1 (IKBKB) S750 Uniprot
O14920 (IKBKB) S756 Uniprot
O43524 (FOXO3) S644 Uniprot
O95999 (BCL10) S134 Uniprot
P01100 (FOS) S308 Uniprot
P04637 (TP53) S362 Uniprot
P04637 (TP53) S366 Uniprot
P19438 (TNFRSF1A) S381 Uniprot
P19838 (NFKB1) S923 Uniprot
P19838-2 (NFKB1) S924 Uniprot
P19838 (NFKB1) S927 Uniprot
P19838-2 (NFKB1) S928 Uniprot
P19838 (NFKB1) S932 Uniprot
P23396 (RPS3) S209 Uniprot
P25963 (NFKBIA) S32 Uniprot
P25963 (NFKBIA) S36 Uniprot
P31946 (YWHAB) S132 Uniprot
P35568 (IRS1) S268 Uniprot
P35568 (IRS1) S270 Uniprot
P35568 (IRS1) S272 Uniprot
P35568 (IRS1) S274 Uniprot
P35568 (IRS1) S307 Uniprot
P35568 (IRS1) S312 Uniprot
P35568 (IRS1) S341 Uniprot
P35568 (IRS1) S345 Uniprot
P35568 (IRS1) S527 Uniprot
P35568 (IRS1) S531 Uniprot
P42771 (CDKN2A) S8 Uniprot
Q01201 (RELB) S472 Uniprot
Q04206 (RELA) S468 Uniprot
Q04206 (RELA) S536 Uniprot
Q04864 (REL) S557 Uniprot
Q13568 (IRF5) S446 Uniprot
Q14164 (IKBKE) S172 Uniprot
Q14653 (IRF3) S396 Uniprot
Q14653 (IRF3) S398 Uniprot
Q14653 (IRF3) S402 Uniprot
Q14653 (IRF3) T404 Uniprot
Q14653 (IRF3) S405 Uniprot
Q15653 (NFKBIB) S19 Uniprot
Q15653-1 (NFKBIB) S23 Uniprot
Q16875 (PFKFB3) S269 Uniprot
Q5S007 (LRRK2) S910 Uniprot
Q5S007 (LRRK2) S935 Uniprot
Q92574 (TSC1) S487 Uniprot
Q92574 (TSC1) S511 Uniprot
Q96CV9 (OPTN) S513 Uniprot
Q99704 (DOK1) S439 Uniprot
Q99704 (DOK1) S443 Uniprot
Q99704 (DOK1) S446 Uniprot
Q99704 (DOK1) S450 Uniprot
Q9BXH1 (BBC3) S10 Uniprot
Q9H3D4 (TP63) S43 Uniprot
Q9H3D4 (TP63) S51 Uniprot
Q9NQC7 (CYLD) S418 Uniprot
Q9NQC7 (CYLD) S422 Uniprot
Q9NQC7 (CYLD) S432 Uniprot
Q9NQC7 (CYLD) S436 Uniprot
Q9NQC7 (CYLD) S439 Uniprot
Q9NQC7 (CYLD) S441 Uniprot
Q9NQC7 (CYLD) S444 Uniprot
Q9Y4G8 (RAPGEF2) S1254 Uniprot
Q9Y6K9-1 (IKBKG) S31 Uniprot
Q9Y6K9 (IKBKG) S43 Uniprot
Q9Y6K9 (IKBKG) S68 Uniprot
Q9Y6K9 (IKBKG) S85 Uniprot
Q9Y6K9-1 (IKBKG) S376 Uniprot
Q9Y6Q9-5 (NCOA3) S857 Uniprot

Research Backgrounds

Function:

Serine kinase that plays an essential role in the NF-kappa-B signaling pathway which is activated by multiple stimuli such as inflammatory cytokines, bacterial or viral products, DNA damages or other cellular stresses. Acts as part of the canonical IKK complex in the conventional pathway of NF-kappa-B activation. Phosphorylates inhibitors of NF-kappa-B on 2 critical serine residues. These modifications allow polyubiquitination of the inhibitors and subsequent degradation by the proteasome. In turn, free NF-kappa-B is translocated into the nucleus and activates the transcription of hundreds of genes involved in immune response, growth control, or protection against apoptosis. In addition to the NF-kappa-B inhibitors, phosphorylates several other components of the signaling pathway including NEMO/IKBKG, NF-kappa-B subunits RELA and NFKB1, as well as IKK-related kinases TBK1 and IKBKE. IKK-related kinase phosphorylations may prevent the overproduction of inflammatory mediators since they exert a negative regulation on canonical IKKs. Phosphorylates FOXO3, mediating the TNF-dependent inactivation of this pro-apoptotic transcription factor. Also phosphorylates other substrates including NCOA3, BCL10 and IRS1. Within the nucleus, acts as an adapter protein for NFKBIA degradation in UV-induced NF-kappa-B activation. Phosphorylates RIPK1 at 'Ser-25' which represses its kinase activity and consequently prevents TNF-mediated RIPK1-dependent cell death (By similarity).

PTMs:

Upon cytokine stimulation, phosphorylated on Ser-177 and Ser-181 by MEKK1 and/or MAP3K14/NIK as well as TBK1 and PRKCZ; which enhances activity. Once activated, autophosphorylates on the C-terminal serine cluster; which decreases activity and prevents prolonged activation of the inflammatory response. Phosphorylated by the IKK-related kinases TBK1 and IKBKE, which is associated with reduced CHUK/IKKA and IKBKB activity and NF-kappa-B-dependent gene transcription. Dephosphorylated at Ser-177 and Ser-181 by PPM1A and PPM1B.

(Microbial infection) Acetylation of Thr-180 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the I-kappa-B pathway.

Ubiquitinated. Monoubiquitination involves TRIM21 that leads to inhibition of Tax-induced NF-kappa-B signaling. According to 'Ser-163' does not serve as a monoubiquitination site. According to ubiquitination on 'Ser-163' modulates phosphorylation on C-terminal serine residues.

(Microbial infection) Monoubiquitination by TRIM21 is disrupted by Yersinia yopJ.

Hydroxylated by PHD1/EGLN2, loss of hydroxylation under hypoxic conditions results in activation of NF-kappa-B.

Subcellular Location:

Cytoplasm. Nucleus. Membrane raft.
Note: Colocalized with DPP4 in membrane rafts.

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

Highly expressed in heart, placenta, skeletal muscle, kidney, pancreas, spleen, thymus, prostate, testis and peripheral blood.

Subunit Structure:

Component of the I-kappa-B-kinase (IKK) core complex consisting of CHUK, IKBKB and IKBKG; probably four alpha/CHUK-beta/IKBKB dimers are associated with four gamma/IKBKG subunits. The IKK core complex seems to associate with regulatory or adapter proteins to form a IKK-signalosome holo-complex. The IKK complex associates with TERF2IP/RAP1, leading to promote IKK-mediated phosphorylation of RELA/p65 (By similarity). Part of a complex composed of NCOA2, NCOA3, CHUK/IKKA, IKBKB, IKBKG and CREBBP. Part of a 70-90 kDa complex at least consisting of CHUK/IKKA, IKBKB, NFKBIA, RELA, ELP1 and MAP3K14. Found in a membrane raft complex, at least composed of BCL10, CARD11, DPP4 and IKBKB. Interacts with SQSTM1 through PRKCZ or PRKCI. Forms an NGF-induced complex with IKBKB, PRKCI and TRAF6 (By similarity). May interact with MAVS/IPS1. Interacts with NALP2. Interacts with TICAM1. Interacts with FAF1; the interaction disrupts the IKK complex formation. Interacts with ATM. Part of a ternary complex consisting of TANK, IKBKB and IKBKG. Interacts with NIBP; the interaction is direct. Interacts with ARRB1 and ARRB2. Interacts with TRIM21. Interacts with NLRC5; prevents IKBKB phosphorylation and kinase activity. Interacts with PDPK1. Interacts with EIF2AK2/PKR. The phosphorylated form interacts with PPM1A and PPM1B. Interacts with ZNF268 isoform 2; the interaction is further increased in a TNF-alpha-dependent manner. Interacts with IKBKE. Interacts with NAA10, leading to NAA10 degradation. Interacts with FOXO3. Interacts with AKAP13. Interacts with IFIT5; the interaction synergizes the recruitment of IKK to MAP3K7 and enhances IKK phosphorylation. Interacts with LRRC14; disrupts IKBKB-IKBKG interaction preventing I-kappa-B-kinase (IKK) core complex formation and leading to a decrease of IKBKB phosphorylation and NF-kappaB activation. Interacts with SASH1. Interacts with ARFIP2.

(Microbial infection) Interacts with Yersinia yopJ.

(Microbial infection) Interacts with vaccinia virus protein B14.

Family&Domains:

The kinase domain is located in the N-terminal region. The leucine zipper is important to allow homo- and hetero-dimerization. At the C-terminal region is located the region responsible for the interaction with NEMO/IKBKG.

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.

Research Fields

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

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

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

· Environmental Information Processing > Signal transduction > NF-kappa B signaling pathway.   (View pathway)

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

· Human Diseases > Drug resistance: Antineoplastic > Antifolate 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 > Epithelial cell signaling in Helicobacter pylori infection.

· Human Diseases > Infectious diseases: Bacterial > Shigellosis.

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

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

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

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

· 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 > Herpes simplex infection.

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

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

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

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

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

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

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

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

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

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

· Organismal Systems > Immune system > Cytosolic DNA-sensing pathway.   (View pathway)

· Organismal Systems > Immune system > IL-17 signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Th1 and Th2 cell differentiation.   (View pathway)

· Organismal Systems > Immune system > Th17 cell differentiation.   (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 > Nervous system > Neurotrophin signaling pathway.   (View pathway)

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

· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.

References

1). Fei H et al. CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway. Front Cell Dev Biol 2021 Aug 4;9:700854. (PubMed: 34422821) [IF=5.201]

2). Xiong Q et al. Nr2e1 ablation impairs liver glucolipid metabolism and induces inflammation, high-fat diets amplify the damage. Biomed Pharmacother 2019 Oct 4;120:109503 (PubMed: 31590127) [IF=4.238]

Application: WB    Species: mouse    Sample: liver

Fig. 5. |Nr2e1 ablation induced hepatic inflammation, the change was aggravated by HFD treatment.The expressions of phosphorylated protein of IKKβ, P65, IκBα in liver samples from the four groups were detected by western blots (n = 3) (5A).

3). Li MY et al. Anti-Inflammatory Effects of Huangqin Decoction on Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice Through Regulation of the Gut Microbiota and Suppression of the Ras-PI3K-Akt-HIF-1α and NF-κB Pathways. Front Pharmacol 2020 Jan 20;10:1552 (PubMed: 32038240) [IF=4.225]

Application: WB    Species: mouse    Sample: colorectal

FIGURE 7 | Effects of Huangqin decoction (HQD) on colonic expression levels of nuclear factor-kappa B (NF-kB) pathway proteins assessed by Western blot in dextran sulfate sodium (DSS)-induced colitis mice (A).

4). Zhang YX et al. Antidepressant-like effects of helicid on a chronic unpredictable mild stress-induced depression rat model: Inhibiting the IKK/IκBα/NF-κB pathway through NCALD to reduce inflammation. Int Immunopharmacol 2021 Apr;93:107165. (PubMed: 33578182) [IF=3.943]

Application: WB    Species: rat    Sample: hippocampus

Fig. 10. The effects of the IKK/IκBα/NF-κB signaling pathway in the hippocampus after AAV-siNCALD. (A) Representative NCALD, IKKβ, p-IKKβ, IкBα, p- IкBα, NF-кB, β-actin, and Lamin B western blot bands. (B), (C), (D), (E), (F), (G), (H) The relative ratios of the grayscale values of NCALD, IKKβ, p-IKKβ, IкBα, pIкBα, and NF-кB to the corresponding β-actin or Lamin B, n = 4. ###P < 0.001, CUMS + saline vs. sham; *P < 0.05; **P < 0.01; ***P < 0.001, CUMS + saline vs. CUMS + AAV-siNCALD. All data are shown as the mean ± SEM.

5). He G et al. Nr2e1 deficiency aggravates insulin resistance and chronic inflammation of visceral adipose tissues in a diet-induced obese mice model. Life Sci 2021 Apr 26;278:119562. (PubMed: 33915130) [IF=3.647]

Application: WB    Species: mice    Sample: adipose tissues

Fig. 5. Nr2e1 deficiency exacerbated inflammation in EAT. Comparison of the epididymal fat weight (A). The ratio of epididymal fat weight to the total body weight (B). The mRNA expression of F4/80 (C). The mRNA and protein expressions of IL-6, IL-1β, TNF-α and MCP-1 (E, F). The protein levels of p-IKKβ/IKKβ, p-P65/P65 (G, H). The data are expressed as means ± SEM. *P < 0.05, **P < 0.01. ns: not significant.

6). Zhao Y et al. Overexpression of endogenous lipoic acid synthase attenuates pulmonary fibrosis induced by crystalline silica in mice. Toxicol Lett 2020 Feb 1 (PubMed: 32017981) [IF=3.569]

Application: WB    Species: mouse    Sample: lungs

Fig. 4. |Effects of enhanced Lias expression on the expression of inflammation markers in mice by western blotting. (A) Levels of NF-κB(P65) and IKK-βprotein in mice lungs in each group, measured by western blotting.

7). Han J et al. Preventive efect of dioscin against monosodium urate‑mediated gouty arthritis through inhibiting infammasome NLRP3 and TLR4/ NF‑κB signaling pathway activation: an in vivo and in vitro study. J Nat Med 2021 Jan;75(1):37-47. (PubMed: 32761488)

Application: WB    Species: human    Sample: synoviocytes

Fig. 4| Dioscin reduced infammation in MSU-treated primary human synoviocytes (HS). Cells were administrated by dioscin (15 ng/mL)for 24 h, and then treated with MSU (100 μg/mL) for 12 h.d The protein expressions of TLR4,MyD88, p-IKKβ, IKKβ, p-p65 in cytoplasm, and NF-κB p65 innuclei were measured utilizing western blotting. GAPDH and Histone H3 were acted as internal references, respectively.

8). Han J et al. Preventive efect of dioscin against monosodium urate‑mediated gouty arthritis through inhibiting infammasome NLRP3 and TLR4/ NF‑κB signaling pathway activation: an in vivo and in vitro study. J Nat Med 2021 Jan;75(1):37-47. (PubMed: 32761488)

9). Huang L et al. Tanshinone IIA ameliorates non-alcoholic fatty liver disease through targeting peroxisome proliferator-activated receptor gamma and toll-like receptor 4. J Int Med Res 2019 Aug 5:300060519859750 (PubMed: 31378113)

Application: WB    Species: rat    Sample:

Figure 3.| Protein and mRNA levels of components of the toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-jB) signalling pathway, levels of liver cell apoptosis and plasma concentrations of interleukin (IL)-6,tumour necrosis factor (TNF)-a and IL-1b in the three study groups: CON (normal feed for 4 months); highfat diet group (HFD for 4 months); and 2 months of HFD followed by 2 months treatment with 10 mg/kg sodium tanshinone IIA sulfonate once daily plus HFD (TAN) group (n ¼ 10 rats per group). TLR4 mRNA levels (a), myeloid differentiation primary response 88 (MyD88) mRNA levels (b), NF-jB mRNA levels (c)and IkB kinase (Ikk-b) mRNA levels (d) were detected by real-time polymerase chain reaction; (e) Western blot analysis to measure levels of TLR4, MyD88, NF-jB, Ikk-b, phospho (p)-IjB-a and phospho (p)-NF-jB(nuclear) proteins in the three study groups. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was the internal loading control and lamin B1 was the nuclear control; (e) Relative levels of each protein according to the Western blot results

10). The Yi-Qi-Bu-Shen recipe attenuates high glucose-induced podocyte injury via the inhibition of IKK-IκBα-NFκB and ERK/P38 MAPK signaling .

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