Journal: Redox Biology
Article Title: Reactive oxygen species promote tubular injury in diabetic nephropathy: The role of the mitochondrial ros-txnip-nlrp3 biological axis
Figure Lengend Snippet: Effects of MitoQ, MSU, TRX siRNA and TXNIP siRNA on TXNIP/NLRP3/IL-1β axis and pro-fibrotic protein expression and apoptosis in HK-2 cells cultured under HG conditions . A : IF showing FN ( Aa-c ) and Collagen I ( Ad-f ) expression in HK-2 cells subjected to HG treatment following MitoQ treatment. B: WB analysis of Caspase-1, cleaved IL-1β and IL-18 expression in HK-2 cells cultured under HG conditions and treated with different concentrations of MitoQ (50,100, 150 nM). C-H: ELISA of Caspase-1 ( C ), IL-1β ( D ), IL-18 ( E ), TGF-β ( F ) activity; TGF-β expression ( G ); and relative apoptosis ( H ) in HK-2 cells subjected to HG conditions and treated with different concentrations of MitoQ (50, 100, 150 nM). I: Luminex analysis of IL-1β, IL-18, FN and Collagen I expression in HK-2 cells subjected to an HG environment and treated with MitoQ, MSU or TXNIP siRNA. J, K: WB and densitometric analysis of NLRP3, IL-1β, IL-18 and FN expression in HK-2 cells exposed to an HG environment and pretreated with MitoQ, TRX siRNA or TXNIP siRNA. Values are the mean ± SE; *P
Article Snippet: 2.13 Analysis of Caspase-1, IL-1β, IL-18 and TGF-β activity Caspase-1 (R & D Systems, USA), IL-1β, IL-18 and TGF-β (Ray Biotech, USA) expression and activity levels in the above cells were detected with the corresponding ELISA Kits, according to the manufacturers’ instructions , .
Techniques: Expressing, Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay, Activity Assay, Luminex