p tak1  (Cell Signaling Technology Inc)

Journal: Chinese Medicine

Article Title: Indole-3-carbinol alleviates allergic skin inflammation via periostin/thymic stromal lymphopoietin suppression in atopic dermatitis

doi: 10.1186/s13020-024-01042-5

Figure Lengend Snippet: Effects of I3C on the activation of MAPKs and NF-κB pathway in DNCB-induced AD skin in NC/Nga mice. A The mRNA expression of TNF-α, IL-1β, IL-6 and IL-4 was determined using RT-qPCR based on GAPDH as the internal control. Total proteins were prepared and western blotted for B p-JNK, JNK, p-p38, p38, C p-TAK1, TAK1, p-IκBα, and IκBα using specific antibodies. Internal control β-actin guided the western blotting for this segment. Densitometric analysis, facilitated by Bio-Rad Quantity One® Software, was applied to the data, illustrating the mean ± S.D. of three independent experiments. ### p < 0.001 vs. the control group; **p < 0.01, and ***p < 0.001 vs. the DNCB-stimulated group

Article Snippet: Primary antibodies against p-p38 (cat no. 9211), p38 (cat no. 9212), c-Jun N-terminal kinase (JNK) (cat no. 9252), p-JNK (cat no. 9251), and p-TAK1 (cat no. 4508) were acquired from Cell Signaling Technology Inc. (Danvers, MA, USA).

Techniques: Activation Assay, Expressing, Quantitative RT-PCR, Control, Western Blot, Software

Journal: Frontiers in Immunology

Article Title: Penta-o-galloyl-beta-d-Glucose (PGG) inhibits inflammation in human rheumatoid arthritis synovial fibroblasts and rat adjuvant-induced arthritis model

doi: 10.3389/fimmu.2022.928436

Figure Lengend Snippet: TAK1 regulates IL-1β-induced IL-6 and IL-8 production in RASF. (A, B) RASFs were pretreated with PGG (5 µM) for 2 hr, followed by IL-1β (10 ng/ml) stimulation for 24 hours. Cytokines array was done as per instruction and developed on X-ray and analyzed with ChemiDoc™ scanning for intensity (A, B, S1 ) IL-6 and IL-8 production was determined in the conditioned media using commercially available ELISA kits. (C, D) RANTES and MMP-1 (E, F) . The values are represented as mean ± SEM of n=4 experiments using different donors. **p<0.01 for IL-1β vs IL-1β+PGG.

Article Snippet: Antibodies against p-TAK1 Thr 184/187 (#4531), p-JNK (#9251) p-JNK (#4511), p-c-Jun (Ser 73 ) (#9164), MyD88 (D80F5) (#4283), CYLD (D6O5O) (#12797) were purchased from Cell Signaling Technology (Beverly, MA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Penta-o-galloyl-beta-d-Glucose (PGG) inhibits inflammation in human rheumatoid arthritis synovial fibroblasts and rat adjuvant-induced arthritis model

doi: 10.3389/fimmu.2022.928436

Figure Lengend Snippet: PGG selectively inhibits phosphorylation of TAK1 at the Thr184/187 site to inhibit its kinase activity. (A) RASF was pretreated with PGG (1-5 µM) for 2 hr, followed by IL-1β stimulation for 30 minutes. Cell lysates were analyzed for MYD88, IRAK-1, IRAK-M, CYLD, A20, TAB1, pTAK1 (Thr 184/187 ), TAK1, TRAF6, I-kBα, p38, pJNK, p-c-Jun, and β-actin. (B) PBMC were pretreated with PGG (5 µM), followed by stimulation with a TLR2 agonist (PamCys3, 1 µg/ml), or TLR4 agonist (lipopolysaccharide, LPS; 1 µg/ml), IL-6 and IL-8 IL-6 and IL-8 production was determined in the conditioned media using commercially available ELISA kits. The results in each figure represent the experiments repeated on four RASF from different donors. (C) RASFs pretreated with PGG (10 μM) and stimulated with IL-1β, immunoprecipitated (IP) with O-GlcNAc or TAB1, and probe for TAK1, TAB1, and p65, also included input for O-GlcNAc to show equal loading. #p<0.05 IL-1b vs. IL-1b+PGG; **p<0.01 for IL-1β vs IL-1β+ PGG.

Article Snippet: Antibodies against p-TAK1 Thr 184/187 (#4531), p-JNK (#9251) p-JNK (#4511), p-c-Jun (Ser 73 ) (#9164), MyD88 (D80F5) (#4283), CYLD (D6O5O) (#12797) were purchased from Cell Signaling Technology (Beverly, MA).

Techniques: Phospho-proteomics, Activity Assay, Enzyme-linked Immunosorbent Assay, Immunoprecipitation

Journal: Frontiers in Immunology

Article Title: Penta-o-galloyl-beta-d-Glucose (PGG) inhibits inflammation in human rheumatoid arthritis synovial fibroblasts and rat adjuvant-induced arthritis model

doi: 10.3389/fimmu.2022.928436

Figure Lengend Snippet: Unique insights from the in silico molecular docking studies of PGG binding in the TAK1-TAB1 complex. (A) Molecular dynamics conformation at 50 ns of PGG in the binding site of TAK1-TAB1 complex. Residue properties surface shown in the Red, Blue and Green represent electronegative, electropositive, and hydrophobic surface area of the binding site. The H-bonds are shown as a dotted line. (B) Inhibition of the TAK-1 in vitro kinase activity by PGG was tested using a kinase assay as per the manufacturer’s instructions. **p<0.01 for No ATP vs ATP. #p<0.05 and ##p<0.01 for ATP vs. PGG.

Article Snippet: Antibodies against p-TAK1 Thr 184/187 (#4531), p-JNK (#9251) p-JNK (#4511), p-c-Jun (Ser 73 ) (#9164), MyD88 (D80F5) (#4283), CYLD (D6O5O) (#12797) were purchased from Cell Signaling Technology (Beverly, MA).

Techniques: In Silico, Binding Assay, Residue, Inhibition, In Vitro, Activity Assay, Kinase Assay

Journal: Frontiers in Immunology

Article Title: Penta-o-galloyl-beta-d-Glucose (PGG) inhibits inflammation in human rheumatoid arthritis synovial fibroblasts and rat adjuvant-induced arthritis model

doi: 10.3389/fimmu.2022.928436

Figure Lengend Snippet: PGG modulates phosphorylation of TAK-1 activity in vivo . (A) H&E staining of rat ankles obtained from normal rats, rats given AIA, and rats were given AIA and Penta-O-galloyl-β-D-glucose (PGG) shows inflammation. In addition, pTAK1 and TAK1 were immunolocalized via IHC staining. (A) Naïve rat ankle stained via H&E shows no inflammation and synoviocytes (SNC), lymphocytes (L), bone (B), and macrophages (M) are present. (B) AIA rat ankle stained by H&E shows high amounts of inflammation including the presence of endothelial cells (EC) which comprise blood vessels (BV) and many synoviocytes (SNC), lymphocytes (L), and macrophages (M). (C) AIA+ PGG rat ankles stained by H&E shows low levels of inflammation and that endothelial cells (EC), blood vessels (BV), synoviocytes (SNC), lymphocytes (L), and macrophages (M) are present. (D) There is no staining for TAK1 in naïve rat ankles. (E) There is minimal staining in synoviocytes (SNC) in AIA rat ankles. (F) There is minimal staining in synoviocytes (SNC) in AIA+PGG rat ankles. (G) There is no staining for p-TAK-1 in naïve rat ankles. (H) There is intense staining for p-TAK-1 in macrophages (M), synoviocytes (SNC), and lymphocytes (L). (I) There is a minimal staining in macrophages (M), synoviocytes (SNC), and lymphocytes (L). NOTE: the same tissue of each treatment was incubated with nonspecific IgG, which showed no expression. (Original magnification × 40). (B) Joint homogenates (30 µg per sample) from naïve, AIA, and PGG treated rats were analyzed for the expression of IRAK-1 (Thr 209 ), pTAK1 (Thr 184/187 ), TAK1, TRAF6, and β-actin.

Article Snippet: Antibodies against p-TAK1 Thr 184/187 (#4531), p-JNK (#9251) p-JNK (#4511), p-c-Jun (Ser 73 ) (#9164), MyD88 (D80F5) (#4283), CYLD (D6O5O) (#12797) were purchased from Cell Signaling Technology (Beverly, MA).

Techniques: Phospho-proteomics, Activity Assay, In Vivo, Staining, Immunohistochemistry, Incubation, Expressing

Journal: Frontiers in Immunology

Article Title: Penta-o-galloyl-beta-d-Glucose (PGG) inhibits inflammation in human rheumatoid arthritis synovial fibroblasts and rat adjuvant-induced arthritis model

doi: 10.3389/fimmu.2022.928436

Figure Lengend Snippet: Schematic figure showing the mechanism of PGG in IL1β signaling. In IL1β signaling, IL-1β activates the signal through its receptor to MyD88, an adaptor protein which further recruits IRAK1-4 and activates TAK1 dependent inflammation. We showed that PGG suppresses IL-1β-induced post translation modification (O-GlcNAC) and downstream inflammatory responses by inhibiting multiple cytokines.

Article Snippet: Antibodies against p-TAK1 Thr 184/187 (#4531), p-JNK (#9251) p-JNK (#4511), p-c-Jun (Ser 73 ) (#9164), MyD88 (D80F5) (#4283), CYLD (D6O5O) (#12797) were purchased from Cell Signaling Technology (Beverly, MA).

Techniques: Post Translation Modification