Fig. 4.

Effects of WSY6 on H₂O₂-induced MAPK activation. (A) PIG1 cells were pretreated with 1 mM H₂O₂ for various times (0, 10, 20, 30, 60 or 120min). Levels of phosphorylated p38 (p-p38), total p38 (p38), phosphorylated ERKs (p-ERKs), total ERKs (ERKs), phosphorylated JNKs (p-JNKs) and total JNKs (JNKs) were examined by Western blots. (Uncropped gel blots are shown in the Supplementary file) (B) Quantitative analysis of p-p38/p38, p-JNK/JNK and p-ERK/ERK. (C) Effects of MAPK inhibitors on cell viability. The cells were pretreated with the p38 inhibitor SB203580, ERK inhibitor U0126 or JNK inhibitor SP600125 for 1 h and then treated in the presence or absence of 1 mM H₂O₂ for 1 h. After treatment, cells were incubated in medium for 24 h. Cell viability was determined with an LDH release assay. (D) PIG1 cells were pretreated with or without EGCG and WSY6 for 1 h, and then treated with 1 mM H₂O₂. Cells were collected at different time points and analyzed by Western blot for the phosphorylation of p38 MAPK. (Uncropped gel blots are shown in the Supplementary file) (E) Quantitative analysis of p-p38 and p38. (n = 3/group, Data were compared using One-Way ANOVA, ns: no significant difference, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).