Fig 4. ADAADi treatment induces apoptosis.

(A). FITC-Annexin V and PI staining in HeLa cells was monitored using FACS in untreated and ADAADi treated cells. (B). Quantitation of the live and apoptotic cells from FACS analysis for HeLa cells. The data is presented as average ± s.d. of three independent experiments. (C). Confocal images obtained after staining untreated and ADAADi treated HeLa cells with acridine orange and ethidium bromide. Green stained cells are viable cells while orange/red cells indicate non-viable or apoptotic cells. (D). The number of viable and apoptotic cells were counted in the untreated and treated HeLa cells. The results are presented as average ± s.d. of two independent experiments. In each experiment, more than 100 cells were counted. (E). The transcript level of CASP3, CASP9, PARP, BAX, Bcl-xl and Bcl2 was measured using qPCR in HeLa cells after treatment with 5 μM ADAADi. GAPDH was used as the internal control. (F). The levels of pro-apoptotic miRNA (underlined in green) and anti-apoptotic miRNA (underlined in red) were analyzed by qPCR in HeLa cells in the absence and presence of ADAADi. In this experiment, RNU6B was used as internal control. The data is presented as average ± s.d. of three independent experiments. Statistical analysis for qPCR experiments was done using Sigma-plot and the star indicates significance at p<0.05.