Fig. 4

IL-1β-induces LPCAT3 via MALT1-c-Myc pathway in chondrocytes. A, B The human chondrocytes were transfected with pCMV-XL4-MALT1 plasmid or empty plasmid (pCMV).48 h post-transfection, the total RNA was isolated and the expression of MALT1 was analyzed by RT-PCR (A). Alternatively, the cells were lysed in 2 × SDS-sample buffer, the proteins were separated by SDS-PAGE and the expression of MALT1 was analyzed by immunoblotting (B). C, D, E The human chondrocytes were transfected with pCMV-XL4-MALT1 plasmid or empty plasmid (pCMV).48 h post-transfection, the cells were treated with IL-1β for 24 h. Later, the total RNA was isolated and the expression of LPCAT3 (C), MMP3 (D), and ADAMTS5 (E) were analyzed by RT-PCR. Data are shown as mean ± SD of 3 independent experiments. ***p < 0.001. F, G The human chondrocytes were transfected with pCMV-XL4-MALT1 plasmid or empty plasmid (pCMV).48 h post-transfection, the cells were lysed in 2 × SDS-sample buffer, the proteins were separated by SDS-PAGE and the expression of c-Myc was analyzed by immunoblotting (F). Alternatively, total RNA was isolated and the expression of c-Myc was analyzed by RT-PCR (G). H, I, J The human chondrocytes were transfected with pCMV-XL4-MALT1 plasmid or empty plasmid (pCMV).48 h post-transfection, the cells were pretreated with c-Myc inhibitors, KJPyr9 (5 μM) or EN4, 5 μM for 1h. Later, the cells were stimulated with IL-1β for 24h. The total RNA was isolated and the expression of LPCAT3 (H), MMP3 (I), and ADAMTS5 (J) were analyzed by RT-PCR. Data are shown as mean ± SD of 3 independent experiments. ***p < 0.001