Figure 3
PLX4720 preferentially increases splicing to produce BimS. (a) Total RNA from Mel-RMu cells treated with PLX4720 (10 μM) for 16 h was subjected to real-time PCR analysis for BimEL, BimL, and BimS mRNA expression. The levels of the expression of individual species before treatment were arbitrarily designated as 1. The data shown are the mean±S.E. of three individual experiments. (b) Total RNA from Mel-RMu cells treated with PLX4720 (10 μM) for 16 h was subjected to BimEL, BimL, and BimS mRNA expression as in a. Left panel: the ratios between the levels of BimS mRNA and BimEL mRNA before and after treatment, respectively, were calculated as (ΔΔCt of BimS/ΔΔCt of BimEL). Right panel: the ratios between the levels of BimS mRNA and BimL mRNA before and after treatment, respectively, were calculated as (ΔΔCt of BimS/ΔΔCt of BimL). The data shown are the mean±S.E. of three individual experiments. (c) Upper panel: total RNA from Mel-RM and Mel-RMu cells treated with SBHA (10 μg/ml) for 16 h were subjected to real-time PCR analysis for Bim mRNA expression. The relative abundance of mRNA expression before treatment was arbitrarily designated as 1. Lower panel: whole cell lysates from cells treated as above were subjected to western blot analysis of Bim and GAPDH (as a loading control). The strong band denoted by the arrowhead is nonspecific, and was associated with the particular batch of the antibody against Bim used in the experiment. The data shown are either representative (lower panel) or the mean±S.E. (upper panel) of three individual experiments. (d) Total RNA from Mel-RM and Mel-RMu cells treated with SBHA (10 μg/ml) for 16 h were subjected to real-time PCR analysis for BimEL, BimL, and BimS mRNA expression. The ratios of BimS mRNA to BimEL and BimL mRNA before and after treatment, respectively, were calculated as in b. The data shown are the mean±S.E. of three individual experiments
