Fig. 6
β-Cyclodextrin-encapsulated resveratrol (Res/CD) induces apoptosis in the hypoxic inner core of HeLa cell spheroids. A HeLa cells grown till confluence were treated with 300 μM CoCl2 and/or 10 μM resveratrol (Res) for 24 h and then stained with annexin-V-FITC. Thereafter, annexin-V-FITC-stained cells were analyzed via flow cytometry. The bar graphs represent the number of apoptotic HeLa cells (mean ± SE, n = 3). p-values were calculated using Student’s t-test. B HeLa cells were treated with 300 μM CoCl2 and/or 10 μM Res for 24 h and stained with 4′, 6-diamidino-2-phenylindole. Fragmented nuclei (indicated using white arrowheads) were observed under a fluorescence microscope. C HeLa spheroid cells were lysed and immunoblotted with an anti-caspase-3 antibody. The levels of cleaved caspase-3 were quantified using densitometry. D To measure caspase-3 activity, HeLa cell spheroids were treated with various reagents mentioned in the Materials and Methods sections. FITC fluorescent intensity (excitation wavelength: 485 nm, emission wavelength: 525 nm) indicates caspase-3 activity. The bar graphs depict caspase-3 activity (mean ± SE, n = 3 ~ 6). p-values were calculated using Student’s t-test. ns = not significant