ARG promotes HCC cells to undergo apoptosis.

<p>(A) Hep G2, SMMC7721 and LO2 cells treated with ARG (0, 1.56, 12.5, 100 μM) for 24 h were double-stained with Annexin V-FITC/PI and analyzed by flow cytometry. Quantification of population rate of early apoptotic cells (Annexin V+/PI− cells, lower right quadrant) and late apoptotic cells (Annexin V+/PI+ cells, upper right quadrants) were shown. Data represent the mean ± SD of triplicate experiments. Significant differences from vehicle-treated control were indicated as *p<0.05, **P<0.01, ***P<0.0001 in each cell line. (B) HepG2 and SMMC7721 cells were treated with ARG at the concentration of 20 μM for 48 h. The nuclei morphology changes were analyzed by Hoechst 33342 staining and the immunofluorescence analysis of cleaved caspase-3 are visualized using the cleaved caspase-3 antibody. The cellular fluorescent changes were observed by fluorescence microscope. At least 200 cells were counted to score the percentage of apoptotic cells which contained both cleaved caspase-3 and condensed and highly fluorescent nuclei fragments in each treatment group. Significant differences from control group were indicated as ***P<0.0001 in each cell line.