Anti tgfβr1

Rabbit polyclonal anti‐GR (Cat: SAB4501309) and mouse monoclonal anti‐α‐smooth muscle actin (Cat: A5228) antibodies were purchased from Sigma (St Louis, MO). Anti–TGFβR1 (Cat: ab31013), anti‐phospho‐smad2 (ab53100), and rabbit polyclonal collagen I (ab34710) were obtained from Abcam (Cambridge, UK). Mouse anti‐β‐catenin (Cat: 610154) and rat anti‐CD31 antibodies (Cat: 550274) were from BD Biosciences (Franklin Lakes, NJ). Fluorescence‐, Alexa Fluor 647–, and rhodamine‐conjugated secondary antibodies were obtained from Jackson ImmunoResearch (West Grove, PA). Reagents included streptozotocin and etomoxir (Sigma).

The proteins were isolated from the frozen kidneys and the concentration was calculated by the Bradford method. Immunoblotting examination was performed as previously described [21 ]. In this study, the primary antibodies used were anti-Ecadherin (1:1000,Abcam,UK), anti-ColIII (11,000, Abcam, UK),anti-caspase1. (1:1000, Abcam,UK), anti-TGFβR1 (1:1000,Abcam,UK), anti-Sirt1 (11,000, Abcam, UK), anti-NLRP3 (1:1000, Abclonal, China), anti-IL-1β (11,000,Abclonal,China), anti-Acetylated-lysine (1:500,Santa Cruz,USA), anti-ASC (1:500, Santa Cruz, USA), anti-TGF-β1 (1500,Santa Cruz,USA), anti-Smad3 (1,1000,CST,USA), and anti-Gapdh (12,000, Proteintech, USA). Protein bands were visualized using the chemiluminescence system (Tanon) for the required time. Quantitative analysis was performed using ImageJ software.