Figure 6.

Effect of IFN-gamma on cardiomyocyte mitochondrial membrane potential. (A) Human cardiomyocytes AC16 were stimulated with 5, 10 or 25ng/ml of IFN-gamma for 48 hours. Then, cells were stained using 1µM TMRM, 400nM of mitotracker DeepRed, 500ng/ml of PI and 1µM of Hoechst 33342 and micrographs were captured in ImageXpress Micro XLS Widefield High-Content Analysis system at 100x magnification. Fluorescence colocalization of TMRE and mitotracker deepred in live cells (PI negative) was used to calculate mitochondrial ΔΨ. Data are reported as the ratio to not-treated cells. Cell viability is the ratio of the number of live cells (PI-negative) and total cells (PI-negative plus PI-positive cells) x 100. n = 3. *p < 0.05. (B) Representative fluorescence microscopy (100x) showing decrease in TMRE fluorescence after incubation with IFN-gamma for 48h.