Supplementary Figure 3 from Ran Is a Potential Therapeutic Target for Cancer Cells with Molecular Changes Associated with Activation of the PI3K/Akt/mTORC1 and Ras/MEK/ERK Pathways

PDF file - 1.6MB, Percentage of sub-G1 phase apoptotic MCF10a and MDA MB231 cells grown in serum-containing medium after Ran silencing. (A-B) Cells were infected with shRan1 and shRan2. Medium was changed to serum-free medium (DMEM) 24hr post-infection. Cells were harvested at 72hr and 96hr post-infection for (A) cell cycle and (B) Western blot analyses. (A) Significantly more sub-G1 phase cells were observed in MDA MB231 breast cancer cells than in immortalized breast epithelial cells 72 and 96 hr post-infection of the shRNAs. (B) More cleaved PARP, caspase 9 and caspase 3 were observed in MDA MB231 than in MCF10A cells upon Ran silencing, suggesting a higher apoptotic response in MDA MB231 than in MCF10a cells. Levels of Ran knockdown were similar in both cell lines. Actin was used as a loading control. (C-D) Cells were infected with shRan1 and shRan2, and harvested 72hr post-infection for (C) cell cycle and (D) Western blot analyses. (C) More apoptotic cells were observed in MDA MB231 than MCF10a cells upon Ran-silencing 72hr post-infection of shRNAs, when these two cell lines were grown in their normal medium. (D) Higher levels of cleaved PARP, caspase 9 and caspase 3 were observed at 72hr post-infection of shRNAs. Results are plotted as histogram showing the mean � SD from three independent experiments. Key; * represents p < 0.05 and ** represents p