Figure 5.

Downregulation of PTBP1 increases the cytoplasmic MCL1 mRNA levels more significantly. Cytoplasmic (C) and nuclear (N) fractions were prepared from PC3 and H1299 cells transfected with either siControl or a mixture of two siPTBP1 for 48 h. Whole-cell protein and RNA extracts (W) were also collected. (a) Protein fractions were analyzed by western blotting to show both the subcellular fractionation quality and that PTBP1 knockdown had no effect on MCL1 protein distribution in cells. GAPDH and Lamin A/C were used as the cytoplasmic and nuclear markers, respectively. (b) RNA fractions were quantified by RT-qPCR using GAPDH as the normalization control. Both relative cytoplasmic and nuclear MCL1 mRNA levels in siControl cells were set as one. Data are shown as mean±S.E.M., n=4. The statistical significance was determined by unpaired student t-test where *P<0.05; **P<0.01; ***P<0.001