Fig. 3.

U73122 combined with apoptosis and necroptosis inhibitors increased Collagen2 and Aggrecan levels in IL‐1β‐treated rat chondrocytes. Chondrocytes pretreated by IL‐1β (20 ng·mL⁻¹ for 36 h) and U73122 (2 μm for 12 h) were treated with Z‐VAD (10 μm for 12 h) or/and Nec‐1 (30 μm for 12 h). Protein (A) and mRNA (B) levels of Collagen2 and Aggrecan were analyzed by Western blotting and RT‐PCR, respectively. β‐Actin was used as the control for Western blotting, and GAPDH was used as the control for RT‐PCR. Values are means and standard deviations, the error bars represent SD. One‐way ANOVA with the Dunnett test was used to calculate P values. These results are representative of at least three independent experiments in each experiment. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.