Fig. 3
From: Inhibition of the MALT1-LPCAT3 axis protects cartilage degeneration and osteoarthritis
Gene silencing or pharmacological inhibition of MALT1 suppresses IL-1β-induced chondrocyte degeneration. A The human chondrocytes were transfected with 200 nM of scrambled RNA (scRNA) or MALT1 siRNA. 48 h post-transfection, the cells were lysed in 2 × SDS-sample buffer, the proteins were separated by SDS-PAGE and the efficiency of knockdown was analyzed by immunoblotting. B, C The MALT1 silenced chondrocytes were treated with IL-1β for 24 h and the cell medium was collected. The total RNA was isolated from the cells and the expression of MMP3 (B) and ADAMTS5 (C) mRNA was analyzed by RT-PCR. The TNFα (D), IL-6 (E), PGE2 (F), and LTB4 (G) levels were analyzed in the medium by ELISA. Data are shown as mean ± SD of 3 independent experiments. ***p < 0.001. H The human chondrocytes were pretreated with vehicle, mepazine (1–10 μM) for 1h, and the cells were stimulated with IL-1β for 24 h. The total RNA was isolated from the cells and the expression of MMP3 (H) and ADAMTS5 (I) mRNA was analyzed by RT-PCR. The TNFα (J), IL-6 (K), PGE2 (L), and LTB4 (M) levels were analyzed in the medium by ELISA. N The human cartilage explants were transfected with 200 nM of scrambled RNA (scRNA) or MALT1 siRNA. 48 h post-transfection, the cells were lysed in 2 × SDS-sample buffer, the proteins were separated by SDS-PAGE and the efficiency of knockdown was analyzed by immunoblotting. O, P The MALT1 silenced human cartilage explants were treated with IL-1β for 24 h and the cell medium was collected. The total RNA was isolated from the cells and the expression of MMP3 (O) and ADAMTS5 (P) mRNA was analyzed by RT-PCR. Q The sGAG in the medium was analyzed by DMMB assay. The PGE2 (R), LTB4 (S), and IL-6 (T) levels in the medium were analyzed by ELISA. U, V The human chondrocytes were transfected with 200 nM of scrambled RNA (scRNA) or MALT1 siRNA. 48 h post-transfection, cells were treated with IL-1β for 24 h, and cell viability (MTT assay) (U), and cell death was analyzed by ELISA (V). In the mepazine group, mepazine (10μM) or vehicle was added 1 h prior to IL-1β treatment. Data are shown as mean ± SD of 3 independent experiments. ***p < 0.001