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. 2019 Oct 16;294(48):18057–18068. doi: 10.1074/jbc.RA119.008925

Figure 5.

Figure 5.

Knockdown of TSG101 diminishes mitophagy and mitochondrial integrity in LPS-treated hearts. A, experimental procedure and biochemical assays for LPS (10 μg/g) treatment in CTRL (TSG101fl/+) and TSG101 KD (MerCreMer-TSG101fl/+) mice. wk, week. B–F Western blots and quantification analysis (B) showing protein levels of TSG101 (C), Parkin (D), PINK1 (E), and LC3-II (F) in hearts of CTRL and KD mice subjected to endotoxin injection for 6 h. GAPDH was used as a loading control for total protein. n = 6 for all groups. * and #, p < 0.05 versus PBS CTRL; &, p < 0.05 versus PBS KD; $, p < 0.05 versus LPS CTRL. G and H, quantification of mtDNA by RT-PCR using primers targeted to mitochondrial cytochrome b (CytB, G) and mitochondrial cytochrome c (CytC, H) in hearts of CTRL and KD mice subjected to endotoxin injection for 6 h. n = 4 for all groups. *, p < 0.05 versus PBS CTRL; #, p < 0.05 versus PBS KD; &, p < 0.05 versus LPS WT. I, levels of mitochondrial hydrogen peroxide (ROS) in hearts of CTRL and KD mice subjected to endotoxin injection for 6 h were measured by Amplex Red assay. n = 6 for all groups. *, p < 0.05 versus PBS CTRL; #, p < 0.05 versus PBS KD; &, p < 0.05 versus LPS CTRL. J, levels of MPO in hearts of CTRL and KD mice subjected to endotoxin injection for 6 h were measured by MPO ELISA kit. n = 4 for all groups. *, p < 0.05 versus PBS CTRL; #, p < 0.05 versus PBS KD; &, p < 0.05 versus LPS CTRL.