Ab131485

For Western blotting, lysates from pancreatic tissue (loading protein = 40 µg) or PSCs (loading protein = 20 µg) were separated through sodium dodecyl sulphate‐polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane. The membrane was incubated with the respective primary antibody overnight at 4°C and then combined with secondary antibodies conjugated to horseradish peroxidase (BA1054 and BA1050; Boster Biological Technology Co., Ltd.). The primary antibodies used were anti‐p65 (ab131485; Abcam Inc), anti‐phosphorylated p65 (ab131109; Abcam Inc), anti‐α‐SMA (bs10196R; Beijing Biosynthesis Biotechnology Co., Ltd), anti‐IκB‐α (D120138; Sangon Biotech Co., Ltd) or anti‐β‐actin (BM0627; Abcam Inc). Detection and image capturing were performed using an enhanced chemiluminescence illuminating system.

M199 medium and fetal calf serum (FCS) were purchased from Gibco/Invitrogen (Karlsruhe, Germany), Penicillin/streptomycin and ß-adrenergic receptor selective antagonists CGP20712A (C231), ICI118.551 (I127) and isoproterenol (I6504) from Sigma-Aldrich (Deisenhofen, Germany). Primary antibodies against the extracellular domain of human RAGE (ab37647), HMGB1 (ab18256), CML (ab27684) and NFkB (ab28856 and ab131485) were from Abcam (Cambridge, UK), against collagen type I from Southern Biotech (1310-01, Southern Biotech, Birmingham, US), against IL-6 (AF 506) and against TNFα (MAB 510) from R&D Systems/Bio-Techne (Wiesbaden, Germany), against F4/80 (#14-4801-82) and against Ly-6G (#14-5931-82) from eBioscience (San Diego, USA). HRP-conjugated secondary antibodies and all other substances used were from Sigma-Aldrich (Deisenhofen, Germany), unless specified otherwise.

Western blot was as described previously [34] (link). We used antibodies against NF-κB (1∶700, t-NF-KB p65 ab131485, p-NF-kB p65, ab28856, Abcam, UK), TXNIP (1∶700, ab86983), NLRP3 (1∶500, ab109314), ASC (1∶700, ab64808), caspase-1 (1∶500, ab1872), IL-1β (1∶1000, ab9722), collagen I (1∶1000, ab34710), collagen III (1∶500, ab7778), and β-actin (1∶1000, sc8432, Santa Cruz, USA). The protein bands were developed by the use of chemiluminescence (Millipore, USA), and quantified by densitometric analysis (Quantity One, Bio-Rad, USA).