Fig. 3.

Effect of β-catenin knockdown on GnRH-induced JNK-c-jun activation. A, LβT2 cells were transfected with either scrambled or β-catenin siRNA for 48 h, serum-starved overnight, and stimulated with 10 nm GnRH or vehicle for 30 min (for JNK) and 1 h (for c-jun). Whole-cell lysates were subjected to a Western blot analysis using β-catenin-, JNK-, phospho-JNK-, c-jun-, and phospho-c-jun-specific antibodies 72 h after transfection. LSD1 was used as a loading control. B, Quantification of Western blot densitometry from three independent experiments, plotted as mean ± sem. Two-way ANOVA. C, Effect of β-catenin or JNK1/2 knockdown on c-jun gene expression. LβT2 cells were transfected with either scrambled, β-catenin, or JNK1/2 siRNA for 48 h, serum starved overnight, and stimulated with 10 nm GnRH or vehicle for 1 h. RNA copy numbers of c-jun were determined by quantitative real time-PCR. Two-way ANOVA with post Bonferroni corrections (n = 3; *, P < 0.05).