GLA induces mitochondrial-mediated apoptosis of UMUC3 cells. (A) UMUC3 cells were treated with various concentrations of GLA (0, 5, 10 and 20 μM) for 24 h. The harvested cells were then stained with Annexin V/PI and flow cytometry was performed to analyze apoptotic rates. (B) The histograms show the percentages of apoptotic cells. (C-E) UMUC3 cells were treated with GLA (0, 20 and 40 μM) for 24 h. Proteins were extracted and western blotting was then performed to analyze the expressions of Bax, Bcl-2, Caspase-9, Cleaved Caspase-9, Caspase-3, Cleaved Caspase-3, PARP and Cleaved PARP. The Bax/Bcl-2 ratio was also calculated as shown in the histogram. Data are expressed as the mean ± S.D. from three independent experiments. *P < 0.05 vs. the control group.