Dlx-2 shRNA prevents metabolic stress-induced loss of ΔΨm and mPT. (A, B) MDA-MB-231 cells were stably transfected with control or Dlx-2 shRNA and treated with GD for 9 h and loaded with 0.5 μM calcein AM and 5 mM CoCl2 for the final 15 min of the incubation. To detect cytoplasmic mitochondrial distribution, 50 nM MitoTracker CMX-ROS were added during calcein loading. Calcein fluorescence was excited at 488 nm and emitted at 515 nm; MitoTracker Red CMX-ROS was excited at 579 nm, and emitted at 599 nm; and the cells were observed using fluorescence microscopy. Representative images of cells from 3 independent experiments were shown (A). The results are expressed as mean ± SE from 15 to 30 cells per treatment group (n = 3) (B). *P < 0.05 versus control; #P < 0.01 versus control shRNA. (C, D) MDA-MB-231 cells stably transfected with control or Dlx-2 shRNA were treated with GD for the indicated times and then treated with 5 mg/ml JC-1 for 15 min. Representative images of cells from 3 independent experiments are shown (C). The results are expressed as mean ± SE from 50 to 100 cells per group (n = 3) (D). *P < 0.05 versus control; #P < 0.05; ##P < 0.01 versus control shRNA.