Figure 9. Cellular localization of BODIPY-DIF-3 in HeLa cells treated with CCCP.

(A) Cells were incubated for 3 days with DMSO (0.2%) or CCCP (10 µM), washed free of the additive, and further incubated for 0.5 h with Hoechst (0.1 µg/ml) and MitoTracker (0.1 µM). Cells were washed free of the additives and observed by using a high-magnification fluorescence microscope. (B) Cells were incubated for 3 days with CCCP (10 µM), washed free of the additive, and further incubated for 0.5 h with BODIPY-DIF-3 (20 µM), Hoechst (0.1 µg/ml), and MitoTracker (0.1 µM). Cells were washed free of the additives and observed by using a high-magnification fluorescence microscope. Three-dimensional (3D) images were constructed, and two representative 2D-projections of the 3D images are shown. Note that CCCP induced mitochondrial swelling (A), which might be slightly enhanced by short-term incubation with BOIDPY-DIF-3 (B).