Fig. 5.

Ripk3-mediated signals are indispensable for persisting synovitis in hTNFtg Ikk2^Ms-KO mice. a Representative histological images of H&E-, TB- and TRAP-stained ankle joint sections of 8-week-old hTNFtg Ikk2^f/f Ripk3^−/− (n = 11) and hTNFtg Ikk2^Ms-KO Ripk3^−/− mice (n = 13) in comparison to hTNFtg Ikk2^Ms-KO and Ikk2^f/f mice. Note the differences in synovium and adjacent tissues in respective areas indicated by the arrows. b Survival rates of Ikk2^Δ/Δ Ripk3^−/− and hTNFtg Ikk2^Δ/Δ Ripk3^−/− SF cultures treated with TNF (T), zVAD (Z), and nec-1 (N) as indicated. c Representative immunodetection of pMlkl, Mlkl and Ripk3 levels in the cell extracts of Ikk2^Δ/Δ Ripk3^−/− and hTNFtg Ikk2^Δ/Δ Ripk3^−/− SFs treated as indicated compared to control Ikk2^Δ/Δ Ripk3^+/+ and hTNFtg Ikk2^Δ/Δ Ripk3^+/+ samples, respectively (n = 4 per genotype, 2 experiments). d Representative immunodetection of Casp8 (C8), Cleaved Casp3 (CC3), Xiap, cFLIP_(s) and IKK2 levels in cell extracts of Ikk2^Δ/Δ Ripk3^−/− and hTNFtg Ikk2^Δ/Δ Ripk3^−/− SFs treated as indicated compared to control Ikk2^f/f Ripk3^−/− and hTNFtg Ikk2^f/f Ripk3^−/− samples, respectively (the non-hTNFtg control cultures are indicated as WT) (n = 4 per genotype, 2 experiments). Scale bar: 500 μm. Data are presented as the mean ± SEM. ***P < 0.001 one-way ANOVA test (b). Unprocessed original scans of blots are shown in Supplementary Fig. 8