Fig. 6

Caspase-3/7 activation in KKU-100 cells following treatment with β-eudesmol, 5-FU, and DOX, and the combination of β-eudesmol and 5-FU or DOX detected by fluorescence microscopy. a KKU-100 cells were treated with 30 μM of β-eudesmol, 30 μM of 5-FU, 0.1 μM of DOX and β-eudesmol (30 μM) combined with 5-FU (30 μM) or DOX (0.1 μM) for 24 h and labeled with CellEvent® Caspase 3/7 Green Detection Reagent and examined under the fluorescence microscope. Left panel, the grey image reveals KKU-100 CCA cells in the field of view. Middle and right panel show apoptotic caspase 3/7 positive cells green fluorescent in the same area of interest. Data are representative of at least 4–5 randomly selected fields’ images with similar results. Morphological changes were evaluated under a microscope. There were shrink and blabbing cells treated with β-eudesmol with or without 5-FU or DOX, indicating apoptosis induction. The scale bar corresponds to 40 μm. b The graph shows the level of caspase-3/7 activity as the percentage of caspase-3 /7 fluorescent cells expression at 24 h. Data are presented as mean ± SD of three independent experiments, triplicate each. * indicates statistically significant difference with each drug alone (paired t-test, p < 0.05). β-Eu= β-Eudesmol, 5-FU= 5-Fluorouracil, DOX= Doxorubicin