Figure 1

Silencing of DNMT3b expression in PC3 cells and validation of cell lines. A. Total RNA as well as protein was extracted from mock (control), and psiDNMT3b transfected PC3 cells and subjected to semi-quantitative RT-PCR analysis and Western blotting. β-actin was used as loading control. B. Affect of DNMT3b silencing on cell proliferation. Cells were grown and their respective rate of proliferation was measured by MTT assay after 24 and 48 hours. Absorbance was measured at 595 nm. C. Apoptosis was studied quantitatively by ethidium bromide and acridine orange staining of control and DNMT3b silenced cells. D. Expression analysis of MCM2. Total protein from mock (control), and psiDNMT3b transfected PC3 cells was subjected to Western blotting to determine expression of MCM2. β-actin was used as loading control. E. Expression analysis of caspase 3. Total protein from mock (control), and psiDNMT3b transfected PC3 cells was subjected to Western blotting to determine expression of caspase 3. β-actin was used as loading control.