Journal: Translational Gastroenterology and Hepatology
Article Title: REGγ deficiency ameliorates hepatic ischemia and reperfusion injury in a mitochondrial p66shc dependent manner in mice
doi: 10.21037/tgh-24-46
Figure Lengend Snippet: REGγ −/− inhibits hepatic mitochondrial injury in mice. In vivo experiment: Transmission electron microscopy, magnification 12,000×, scale bar 500 nm (A), Mitochondrial average diameter analysis (B); WB of p-DRP/DRP (C), gray value analysis (D); WB of Cytochrome C (E), gray value analysis (F); n=5 per group, *, P<0.05 vs. WT-Sham group; # , P<0.05 vs. WT-I/R group. In vitro experiments: JC-1 fluorescence staining 20× and MFI analysis (G,H). ROS fluorescence staining was performed 20×, and MFI analysis (I,J). n=5 per group, *, P<0.05 vs. WT-control group; # , P<0.05 vs. WT-H/R group. I/R, ischemia and reperfusion; WT, wild type; REG, the 11S proteasome regulatory complex; DRP, dynamin related protein 1; ROS, reactive oxygen species; WB, Western blot; MFI, mean fluorescence intensity; H/R, hypoxia and reoxygenation.
Article Snippet: Primary antibodies as following: 78–82 Kda hosphor-DRP1 Rabbit 1:1000, CST (Shanghai, China); 83 Kda DRP1 Rabbit 1:1000, Abcam (Shanghai, China); 61/55 Kda hosphor-NF-κB p65 Rabbit 1:1000, Bioss (Beijing, China); 65 Kda NF-κB p65 Rabbit 1:2000, Bioss; 36 Kda p-IκBα Rabbit 1:2000, CST; 36 Kda IκBα Rabbit 1:2000, CST; 35 Kda Caspase-3 Rabbit 1:1000, CST; 17/19 Kda Cleaved Caspase-3 Rabbit 1:1000, CST; 21 Kda BCL2-associated X protein (BAX) Rabbit 1:1000, Proteintech (Wuhan, China); 26 Kda B-cell lymphoma-2 (BCL-2) Rabbit 1:1000, Proteintech; 42 Kda β-actin Mouse 1:5000, Proteintech; 63/67–75 Kda p66shc Rabbit 1:1000, Proteintech; 63/55 Kda p-ser36 p66shc Mouse 1:1000, Abcam; 36 Kda GAPDH Rabbit 1:5000, Affinity (Nanjing, China); 12–15 Kda Cytochrome C Mouse 1:5000, Proteintech; 17–18 Kda COXIV Rabbit 1:5000, Proteintech; 35 REGγ Rabbit 1:2000, Proteintech.
Techniques: In Vivo, Transmission Assay, Electron Microscopy, In Vitro, Fluorescence, Staining, Control, Western Blot
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