Effect of genipin on apoptosis.

<p>The effect of genipin on apoptosis was investigated by a caspase 3/7 assay, which indicates the apoptotic activity. (A) The caspase 3/7 assay demonstrated that the effects of genipin on apoptosis were significantly different in iSLK-puro and iSLK-BAC16 cells. Apoptosis was slightly suppressed in iSLK-BAC16 cells by the genipin treatment at both concentrations. (B) Western blot assays using antibodies to apoptosis signaling-related proteins, such as anti-PARP, anti-BAX, anti-Bcl-2, and anti-cytochrome C antibodies, were conducted to determine the effects of genipin on the induction of apoptosis in iSLK-puro and iSLK-BAC16 cells. (C) Intensities of bands in Western blot assay data were quantitated using GelQuant.NET program. Resultant band intensities standing for PARP, BAX, Bcl-2 and cytochrome C were normalized to the band intensity standing for GAPDH. After that, normalized intensities of bands from genipin treatments at different concentrations were compared with those from the 0 μM genipin treatments. (D) The cell cycle assay was conducted using the Muse^TM Cell Cycle Kit to determine whether the 48-h treatments of genipin at different concentrations made any effect on cell cycle progresses of iSLK-puro and iSLK-BAC16 cells. Results are averages of values from five independent experiments. Standard errors of the means are indicated with the T bars. *, # and ^ stand for statistical significant between two corresponding groups in ANOVA analysis.