Antibody against β actin

Fetal bovine serum (FBS) was purchased from Thermo Fisher Scientific (Shanghai, China). ISO was procured from Sigma Industry Co., Ltd. (St Louis, MO, USA). In addition, dual anti-sterilization solution (penicillin/streptomycin solution), 0.25% trypsin-EDTA, phosphate-buffered saline (PBS), and Dulbecco’s modified Eagle’s medium (DMEM) were all purchased from Meilun Biotech (Dalian, China). MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) was purchased from Beyotime Biotechnology (Shanghai, China). Dimethyl sulfoxide (DMSO) was purchased from Meilun Biotech (Dalian, China).
Antibody against β-actin was purchased from Servicebio (Wuhan, China), and the caspase-3, caspase-1, PARP, and cleaved PARP antibodies were obtained from Biosystems Dogs (Shenyang, China). Other antibodies were all purchased from Beyotime Biotechnology (Shanghai, China).

Proteins were extracted by using RIPA buffer, and protein expression levels were examined by Western blot as described previously. It should be noted that for the fecal proteins, approximately 100 mg of feces was collected and lysed with RIPA lysis buffer (in a water bath at 37°C for 30 min). After electrophoresis, the proteins were transferred onto nitrocellulose membranes, which were incubated with primary antibodies, followed by incubation with HRP-conjugated secondary antibodies. The immunodetected proteins were visualized using an ECL assay kit (Millipore, Burlington, USA). The antibody against β-actin (Cat#: GB11001) was obtained from Servicebio (Wuhan, China; 1:2000 dilution). The antibody against GSDMD (#sc81868) was purchased from Santa Cruz (Dallas, USA; 1:500 dilution). The antibodies against IL-1β (#A1112) and NLRP3 (#A5652) were purchased from ABclonal (Cambridge, USA; 1:1000 dilution). The antibodies against Caspase-1 (#22915-1-AP), ZO-1 (#21773-1-AP), Occludin (#27260-1-AP) and Claudin-1 (#13050-1-AP) were obtained from Proteintech (Chicago, USA; 1:1000 dilution).

Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were produced by Gibco and purchased from Thermo Fisher Scientific (Waltham, MA, USA). Complete Mycobacterium medium was purchased from Shanghai Gene-Optimal Science & Technology Co., Ltd (Shanghai, China).
Opti-MEM I reduced serum medium was produced by Gibco and purchased from Thermo Fisher Scientific. FuGENE^® HD Transfection Reagent was obtained from Promega (Beijing) Biotech Co., Ltd (Beijing, China). Antibodies against DUSP10 (# ab228987) and LC3-I/II (# ab128025) were purchased from Abcam (Cambridge, UK). Antibodies against Beclin1 (# D160120) and Atg5 (# D121650) were procured from Sangon Biotech (Shanghai) Co., Ltd (Shanghai, China). Antibodies against Atg7 (# AA820) and p-Erk1/2 (# AF5818) were purchased from Beyotime Biotech Inc. (Shanghai, China). Antibody against β-actin (# P60710) was purchased from Servicebio (Wuhan, China). Horseradish peroxidase (HRP)-labeled goat anti-rabbit IgG (# SSA004) was purchased from SinoBiological (Beijing, China). Anti-Rabbit IgG (whole molecule)-Gold (# G7402) was purchased from Sigma–Aldrich (Shanghai) Trading Co. Ltd (Shanghai, China). ERK1/2 signaling inhibitor PD98059 (# HY-12028) was procured from MCE (MedChem Express, Monmouth Junction, NJ, USA).