Short hairpin RNA-mediated knockdown of nuclear factor erythroid 2-like 3 exhibits tumor-suppressing effects in hepatocellular carcinoma cells

Figure 3 Role of nuclear factor erythroid 2-like 3 in apoptosis, clone formation, and proliferation of hepatocellular carcinoma cells. A and B: Apoptotic cells were stained with Annexin V-APC and measured using flow cytometry. The abscissa represents red fluorescence (RED-R-HLog), and the ordinate represents green fluorescence (GRN-B-HLog) and cell count; C and D: Cell clones were stained with Giemsa and photographed with a digital camera. The number of clones was accurately calculated and statistically analyzed; E and F: Successfully infected cells were green fluorescent protein positive and cell images were taken with a Celigo cytometer for a continuous 5 d. Cell count-fold represents cell count at each time point relative to the average of day 1; G and H: OD values were measured at 490 nm after the treatment of MTT. Cell growth curves were plotted based on OD 490-fold at different time point. Data shown are the mean ± SEM. Student’s t test was used to analyze significant differences, ^aP < 0.05, ^bP < 0.01, ^cP < 0.001 vs shCtrl. shCtrl: Negative control cells; shNFE2L3: Nuclear factor erythroid 2-like 3 short hairpin RNA.