Figure 2
ROS generation and decreased ΔΨm in poly(I:C)-transfected RCC cells. (a) RCC cells were transfected with poly(I:C) (SKRC-1, 500 ng/ml; SKRC-44, 50 ng/ml) and cultured for 24 h in the presence or absence of NAC (10 mM). Treated cells were stained with carboxy-H2DCFDA. Numbers represent the mean fluorescence intensity (MFI) of cells in each subset. Similar results were obtained from three independent experiments. (b) Cells were then stained with Annexin V-FITC and PI and analyzed by flow cytometry. Numbers represent the percentages of cells in each subset. (c) RCC cells were transfected with poly(I:C) (SKRC-1, 1,000 ng/ml; SKRC-44, 500 ng/ml) and cultured for 24 h in the presence or absence of NAC (10 mM). Similar results were obtained from three independent experiments. RCC cells were then examined for ΔΨm levels by flow cytometry using Alexa Fluor 488-conjugated MitoProbe™ DiOC2(3) (8 nM). Numbers represent the percentages of positive cells with ΔΨmhigh. Similar results were obtained from three independent experiments. (d) RCC cells were then stained with Alexa Fluor 488-conjugated DiOC2(3) (8 nM) and Annexin V-APC and analyzed by flow cytometry. Numbers represent the percentage of positive cells in each subset. Similar results were obtained from two independent experiments. pIC-TF, poly(I:C) transfection.