Journal: Translational Andrology and Urology
Article Title: Suppression of NLRP3 inflammasome activation by astragaloside IV via promotion of mitophagy to ameliorate radiation-induced renal injury in mice
doi: 10.21037/tau-23-323
Figure Lengend Snippet: Animal treatment and the effects of radiation on renal function, ROS level, and expression of Cytc. (A) The timeline shows the time points for drug treatment and radiation. (B) SCr (a), BUN (b), and UA (c) levels in different groups of mice are shown. (C) ROS levels in different groups of mouse kidneys were evaluated by dihydroethidium staining (400×). (D) Western blot expression results and the quantification of the levels of Cytc in cytoplasm (a,c) and mitochondria (b,d) in the kidneys of each group, with normalization to GAPDH and COX-IV respectively. The data are expressed as the mean ± standard error of the mean (n=3; **, P<0.01 and ***, P<0.001 vs. the control group; # , P<0.05, ## , P<0.01 and ### , P<0.001 vs. the DMSO group). ROS, reactive oxygen species; Cytc, cytochrome C; AS-IV, astragaloside IV; CsA, cyclosporin A; SCr, creatinine; BUN, blood urea nitrogen; UA, uric acid; DMSO, dimethyl sulfoxide; IR, irradiation; c-Cyt c, Cytc in the cytosol; COX-IV, Cytc oxidase IV; mito-Cyt c, Cytc in the mitochondria.
Article Snippet: This was followed by blocking in 5% skimmed milk in tris-buffered saline with Tween20 (TBST) at RT for 1 h and overnight incubation with primary antibodies against c-Cytc (bs-0013R; Bioss Antibodies), NLRP3, cleaved caspase-1 (bs-10442R; Bioss Antibodies), IL-1β (bs-0812R; Bioss), Bax (A00183; Boster Bio), B-cell lymphoma-2 (Bcl-2; BA0412; Boster Bio), prostacyclin 62 (P62; bs-2951R; Bioss Antibodies), parkin (bs-23687R; Bioss Antibodies), LC3, PINK1 (23274-1-AP; Proteintech), cleaved caspase-3, cleaved caspase-9 (20750S; Cell Signaling Technology, Danvers, MA, USA), Cytc oxidase IV (COX-IV; 11242-1-AP; Proteintech), β-actin (bs-0061R; Bioss Antibodies), and GAPDH (K200057M, Solarbio) at 4 °C, with subsequent incubation in corresponding HRP-conjugated secondary antibodies at RT for 2 hours.
Techniques: Expressing, Staining, Western Blot, Control, Irradiation