Ab260017

Dulbecco’s modified eagle medium (DMEM) and fetal bovine serum (FBS) were purchased from life technologies (Kalamazoo, MI, USA). BGC823 cells were employed to be in vitro model to study the function of Rabeprazole [11 ]. The human gastric epithelial cell BGC823 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in DMEM supplemented with 10% FBS. The cells were maintained at 37 °C in a humidified 5% CO₂ incubator. Pierce™ BCA protein assay Kit and PageRuler™ Prestained Protein Ladder were purchased from thermo fisher, trizol was from invitrogen (Invitrogen, Thermo Fisher Scientific). All-in-one™ first-strand cDNA synthesis kit and All-in-one™ qPCR mix were from Genecopoeia™ (Rockville, MD, USA). Rabeprazole (S4845) was purchased from Selleck. LPS (BS904) was purchased from Biosharp. Other chemical reagents were from Sigma. Antibodies were purchased from Abcam: NLRP3(Abcam; ab260017, 1:2000 for WB); IL-18(Abcam; ab235697, 1:1000 for WB); IL-1β (Abcam; ab216995, 1:1000 for WB); ASC (Abcam; ab151700, 1:2000 for WB); GSDMD (Abcam; ab210070, 1:2000 for WB); Human IL-18 ELISA Kit (ab215539) and Human IL-1 β ELISA Kit (ab217608) were purchased from Abcam. β-actin (AC038) purchased from Abclonal. For treatment, BGC823 cells were stimulated for Rabeprazole (10uM) for 1 h, following by LPS (500 ng/mL) treatment for further 48 h.

According to the instructions, cerulein-treated AR42J acinar cells with RIPA buffer (Beyotime) and 30 μg denatured proteins were subjected to detect protein expression. Membrane with protein attached was incubated with primary antibodies against TRAF6 (1:1000, AB137452, Abcam), TLR9 (1 : 200, ab134368, Abcam), NLRP3 (1 : 1000, ab260017, Abcam), caspase-1 (1 : 1000, ab179515, Abcam), ASC (1 : 1000, ab283684, Abcam), and GSDMD (1 : 1000, ab210070, Abcam). The loading control was GAPDH (1 : 10 000, ab181602, Abcam). Band analysis software is ImageJ (version 1.8.0_172).