Fig. 5: The TAK1/IKK2 axis regulates TNF arthritogenicity in TNF-mediated arthritis. | Cell Death & Disease

Fig. 5: The TAK1/IKK2 axis regulates TNF arthritogenicity in TNF-mediated arthritis.

From: Cyld restrains the hyperactivation of synovial fibroblasts in inflammatory arthritis by regulating the TAK1/IKK2 signaling axis

Fig. 5

a Detection of Tak1 ubiquitination by immunoassay of Cyldf/f and CyldM-Δ9/Δ9 SFs stimulated with TNF (0–60 min), followed by immunoprecipitation of lysates with anti-Tak1 and immunoblot analysis with anti-K63-Ub, and then re-probing with anti-Tak1 and anti-Tab1. (Lower panel) Immunoblot analysis of corresponding whole cell lysates (input) with anti-Cyld, anti-Tak1 and anti-Tab1 (n = 3). b Detection of Tak1 phosphorylation by immunoassay of Cyldf/f and CyldM-Δ9/Δ9 SFs stimulated with TNF (0–30 min), followed by immunoprecipitation of lysates with anti-Tab1 and immunoblot analysis with anti-pTak1. c Histological evaluation of 10-week-old hTNFTg Ikk2f/f Cyldf/f, hTNFTg Ikk2M-HetCyldΜ-Δ9/Δ9 and hTNFTg Ikk2M-KOCyldΜ-Δ9/Δ9 mice; Scale bar: 500 μm. d Survival rates of Cyldf/f and CyldM-Δ9/Δ9 SFs treated either with 50 ng/ml TNF or combinations of TNF with zVAD or Nec1 as indicated (T + Z: TNF 50 ng/ml + zVAD, T + N1s: TNF 50 ng/ml+ Nec1s) in the absence (Black bars) or presence (Gray bars) of ΙΚΚ2 inhibitor ML120b, s (n = 3–5). Data are presented as the mean ± SEM *p < 0.05, **p < 0.01, and ****p < 0.0001 by one-way ANOVA (Bonferroni correction) (c) or two-tailed Student’s t-test (d).

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