Fig. 2

Inflammatory response causes mitochondrial dysfunction in RAW 264.7 cells during macrophages polarization. The RAW 264.7 cells were treated with LPS (100 ng/ml) or LPS + IFN-γ (20 ng/ml) for 24 h. (A) The M1 markers (iNOS and TNF-α) and the M2 markers (CD206 and Arg-1) were measured by western blotting. (B) The MMP was determined using a JC-1 probe (immunofluorescence straining; scale bar: 10 μm). (C) The marker of mitochondria (TOM 20) was detected by western blotting. (D) The RAW 264.7 cells were immunostained with 100 nM Mito-Tracker (green) plus 100 nM Mito-Tracker Red CMXRos (red). Quantification of depolarization (damaged) mitochondria. Scale bar: 10 μm. (E) Quantification of mitochondrial ROS levels were determined by MitoSOX (red). Scale bar: 10 μm. Meanwhile, (F) The level of GSH/GSSG and the level of SOD were measured in RAW 264.7 cells pretreated with LPS or LPS + IFN-γ for 24 h. ^*P < 0.05, ^**P < 0.05 vs the control group