Figure 3
The ERK signaling pathway promotes RNF126 expression. (a and b) Western blot analysis of ERK1/2 and phosphorylated (p)-ERK1/2 in control and RNF126-depleted MDA-MB-231 (a) and A549 cells (b). (c and d) Western blot analysis of RNF126, ERK1/2 and phosphorylated-ERK1/2 in vehicle (dimethyl sulfoxide (DMSO))- or MEK inhibitor U0126- (10 μm) treated MDA-MB-231 (c) and A549 cells (d). Results of the densitometric analysis of bands in western blots are presented. (e and f) mRNA levels of RNF126 in vehicle (DMSO)- or MEK inhibitor U0126- (10 μm) treated MDA-MB-231 (e) and A549 cells (f) were analyzed by real-time PCR. Error bars indicate the s.d. (n=3). The data were analyzed by a t-test. **P<0.01. (g) RNF126 promoter activity was analyzed in MDA-MB-231 cells treated with or without U0126 (10 μm). Error bars indicate the s.d. (n=3). The data were analyzed by a t-test. **P<0.01. (h) ChIP real-time PCR of the RNF126 promoter using anti-ELK-1 or control immunoglobulin G (IgG) in MDA-MB-231 cells (left) and A549 cells (right) treated with or without U0126 (10 μm). Error bars indicate the s.d. (n=3). The data were analyzed by a t-test. **P<0.01. The data shown in (a–h) are representative of three independent experiments with similar results.
