Research Paper Volume 12, Issue 2 pp 1114—1127

Figure 3. The overexpression of Romo1 promoted the accumulation of ROS and results in mitochondrial dysfunction in macrophages. (A) The control and the Romo1-overexpressed macrophages were analyzed by immunofluorescence with DCF-DA and MitoSOX staining to detect the levels of cytoplasmic and mitochondrial ROS. The ROS+ cells were counted under microscopy and statistically analyzed. (B) The levels of DHE in control and Romo1-overexpressed macrophages (with or without treatment of 5mM NAC) were analyzed by flow cytometry. (C) The mean fluorescence intensity (MFI) of DHE in flow cytometry was respectively quantified and statistically analyzed. (D) The statistical analysis of the mtDNA copy number in control and Romo1-overexpressed (with or without treatment of 5mM NAC) macrophages. (E) The statistical analysis of the oxygen uptake rate in control and Romo1-overexpressed (with or without treatment of 5mM NAC) macrophages. (F) The expression of the indicated proteins was examined by western blotting in control and Romo1-overexpressed (with or without treatment of 5mM NAC) macrophages. Data are representative of at least three independent experiments and are presented as mean ± SD. ns, not significant; *, P < 0.05; **, P < 0.01.