Fig. 2.

Role of mitochondria in CO-induced HIF-1α expression. (A) Mitochondria-deficient (ρ°) and WT mφs were characterized by their ability to generate ROS by 2′,7′-dichlorodihydrofluorescein diacetate (DCF) fluorescence. (Upper) WT. (Lower) ρ°. Cells were preloaded with DCF ± CO, and fluorescence was determined by FACS (filled, 0 min; unfilled dashed line, 5 min; unfilled bold line, 60 min). (B) Western blot showing kinetics of HIF-1α protein in ρ° cells exposed to CO; α-tubulin is shown as a loading control. (C) Immunofluorescent staining for HIF-1α (red, denoted by arrows) in cultured WT mφs (left-air and right-CO; Upper) and ρ° cells (left-air and right-CO; Lower). Images shown are representative of at least 10 fields of view. Results shown represent one of three independent experiments. (Scale bar: 1 μm.)