A1170

Western blot was performed as described previously. 24 (link) Specific proteins were detected using the following primary antibodies: NLRP3 (no. ab214185, 1:1000, Abcam), ASC (no. A1170, 1:1000, ABclonal), caspase-1 (no. ab238972, 1:1000, Abcam), IL-1β (no. ab9722, 1:1000, Abcam) and GSDMD (no. ab219800, 1:1000, Abcam). Horseradish peroxidase-conjugated secondary antibodies (ZB-2301, ZB-2305, 1:1000, ZSGB) were incubated with the membrane and the antibody complexes were detected by Imaging System (Bio-Rad, Hercules, CA, USA). β-Actin (no.TA-09, 1:1000, ZSGB) was used as the control to detect equal protein loading.

WB was performed according to our previous study method [17 (link), 35 (link)]. We used the following primary antibodies to perform the WB analyses: rabbit monoclonal anti-NF-κB p65 (D14E12) XP® antibody (1:1000, #8242, Cell Signaling Technology); rabbit polyclonal anti-Nrf2 (L593) antibody (1:500, BS1258, Bioworld); mouse monoclonal anti-Cryopyrin (NLRP3) (6F12) antibody (1:1000, sc-134306, Santa Cruz Biotechnology); rabbit polyclonal anti-PYCARD (ASC) antibody (1:500, A1170, Abclonal); goat polyclonal anti-caspase-1 p20 (M-19) antibody(1:1000, sc-1218, Santa Cruz Biotechnology); rabbit polyclonal anti-IL-1β (H-153) antibody (1: 1000, sc-7884, Santa Cruz Biotechnology); and rabbit polyclonal anti-IL-18 (H-173) antibody (1:1000, sc-7954, Santa Cruz Biotechnology), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:1000, Cell Signaling Technology) was used as an internal reference. Blot bands were quantified via densitometry with ImageJ software (National Institutes of Health, Baltimore, MD, USA), and protein levels were expressed as the ratio of values of the detected protein bands to that of GAPDH bands.