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. 2021 Jul 16;12:675534. doi: 10.3389/fimmu.2021.675534

Figure 5.

Figure 5

Effects of ATG administration on the MSU crystal-induced production of intracellular total ROS, mitochondrial ROS and mitochondrial membrane potential. (A, B, D, E) J774A.1 cells were pretreated with ATG (5 μM) for 1 h, primed with LPS for 1 h and then treated with MSU crystals (50 μg/ml) for 9 h. *P < 0.05 vs. without MSU crystals treatment; #P < 0.05 vs. MSU crystals treatment +vehicle. (C, F) J774A.1 cells were either transfected with scRNA or siTTP for 48 h, treated with 5 μM ATG for 1 h, primed with LPS for 1 h, and then stimulated with MSU crystals (50 μg/ml) for 9 h. *P < 0.05 vs. MSU crystals treatment + ATG + scRNA; #P < 0.05 vs. MSU crystals treatment + scRNA. (A, B) Cells were stained with DHE fluorescent probe, then the fluorescence intensity of DHE was measured by FACS or images were captured by Laser confocal microscope. Scale bar: 10 μm. (C) The fluorescence intensity of DCFH was measured by FACS. (D) Cells were stained with MitoSOX and then FACS was used to detect the fluorescence intensity of MitoSOX. (E) Cells were stained with MitoSOX and Hoechst 33342. Laser confocal microscope is used for image capture and fluorescence intensity analysis. Blue shows nuclei stained with Hoechst 33342. Scale bar: 20 μm. (F) FACS was used to detect the fluorescence intensity of MitoSOX. All the data are expressed as the means ± SEM from n=3 independent experiments.