Fig. 3: Cyld-DUB-deficient SFs exhibit enhanced expression of arthritic mediators. | Cell Death & Disease

Fig. 3: Cyld-DUB-deficient SFs exhibit enhanced expression of arthritic mediators.

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

Fig. 3

a Flow cytometric analysis of SF subtypes (Τhy1-, Lining SFs; Thy1+, Sublining SFs) and myeloid infiltrates in the ankle joints of hTNFTg Cyldf/f and hTNFTg CyldM-Δ9/Δ9 4-week-old mice compared to WT controls. b Cyld expression in SF cultures derived from indicated genotypes. Extracts from Cyld-deficient MEFs served as control. c Relative gene expression of arthritic mediators Il1b, Il6, Mmp3, Mmp9, Mmp13, and Timp1 (normalized to b2m levels) in SFs of Cyld-proficient and Cyld DUB-deficient SFs upon TNF (20 ng/ml, 24 h) (n = 4). d Relative gene expression of arthritic mediators Il1b, Il6, Mmp3, Mmp13 and Timp1 (normalized to b2m levels) in SFs of hTNFTg Cyld-proficient and Cyld DUB-deficient SFs. e Representative gelatin-based zymography utilizing supernatants of SF cultures derived from hTNFTg Cyld-proficient and Cyld DUB-deficient SFs. f hTNF protein levels in cultures of hTNFTg Cyld-proficient and Cyld DUB-deficient SFs (n = 3–6). g Flow cytometric detection of Vcam-1 and Icam-1 levels in cultured hTNFTg Cyldf/f (black), hTNFTg CyldM-Δ9/Δ9 (gray) (upper panel) and representative FACS plots (n = 8, 2 experiments). Data are presented as the mean ± SEM (or mean ± SD for f) *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 by two-way ANOVA (Bonferroni correction) (a) one-way ANOVA (Bonferroni correction) (c) or unpaired t test with Welch’s correction (d, g).

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