Increased apoptosis upon TGS of
c-Myc shRNA constructs were transfected in HepG2 cells and post 5 days, apoptosis (subG1) was evaluated by flow cytometry. Increase in apoptosis was found to be concordant with the strength of promoter/enhancer based construct driving shRNA transcription and corroborated with the MTT data. Since the fixed amount of each construct was utilized, maximum percentage of cells in subG1 phase was observed through AFPEn–Pr + 2 – myc construct whereas it was the lowest in the case of AFPPr + 2 – myc. No significant apoptosis was seen by the scrambled c-Myc shRNA under CMV promoter (CMVPr – myc Scr). In addition, c-Myc suppressed cells showed reduced S and G2M phase. (B) In the case of Chang Liver cell line, only CMVPr – myc clearly shows significant population of apoptotic cells due to its non-specific nature.