Ficus induces apoptosis in HeLa through mitochondrial dependent pathway.

<p>(<b>A</b>) Representative FACS pictograms of cells treated with FR<b>_aq</b>(0–80 µg/ml) are shown. Percent of annexin V-positive (early-apoptotic cells, lower right quadrant) and Annexin V/PI-double-positive cells (late-apoptotic cells, upper right quadrant) are indicated. (<b>B</b>) Flow cytometric analysis of the rapid calcium release in HeLa cells after treatment with FR<b>_aq</b>(0–80 µg/ml) has been shown. Ionomycin was used as a positive control. The data represents mean ± SD of three independent experiments. (<b>C</b>) FACS analysis following JC-1 staining of HeLa showed alteration of the mitochondrial membrane potential after FR<b>_aq</b>(0–80 µg/ml) treatment compared to untreated control cells. The data represents mean ± SD of three independent experiments. (<b>D</b>) Western blot shows the expression of cytochrome c from cytosolic fraction. Tubulin was used as a loading control. (<b>E</b>) Total protein was isolated and analysed for expression of p53 and caspase 3 by immunoblotting. Tubulin was used as a loading control. (<b>F and G</b>) Densitometric analysis of the western blot showing fold change in protein levels. The bands were quantified by using ImageJ 1.44p (Wayne Rasband, National Institutes of Health, USA, <a href="http://imagej.nih.gov/ij" target="_blank">http://imagej.nih.gov/ij</a>).