Ab108335

Radioimmunoprecipitation assay (RIPA; Beyotime, China) buffer was applied for protein extraction from tissues. The supernatants were run on 8%–12% acrylamide gels via SDS-PAGE and then transferred onto polyvinylidene difluoride membranes (Millipore, USA). Antibodies against GPX4 (#P36969; RayBiotech, USA; 1:1000), NLRP7 (#AB117732; Abcam, USA; 1:1000), CASP6 (#AB108335; Abcam; 1:1000), NLRP6 (#P59044; RayBiotech; 1:1000), IL-6 (#AB271042; Abcam; 1:2000), CASP5 (#AB40887; Abcam; 1:3000), CASP4 (#AB238124; Abcam; 1:1500), TIRAP (#P58753; RayBiotech; 1:1000), and GAPDH (#60004-1-Ig; Proteintech, China; 1:20000) and HRP-conjugated secondary antibodies (#ab97080 or ab47827; Abcam; 1:2000) were employed for western blot. The chemiluminescence western blot detection system (Bio-Rad, USA) was utilized for protein detection.

Western blotting was carried out according to standard protocols (Ma et al., 2019 (link); Yang et al., 2020 (link)). Antibodies used in the western blot are listed as follows: anti-PAM (Abcam, ab237488), anti-SLC16A3 (Abcam, ab234728), anti-Stanniocalcin 1 (Abcam, ab124891), anti-CLDN9 (Proteintech, 16196-1-AP), anti-HDAC4 (Proteintech, 17449-1-AP), anti-KDELR3 (Proteintech, 27632-1-AP), anti-Versican (Abcam, ab19345), anti-triosephosphate isomerase (Abcam, ab170894), anti-CASP6 (Abcam, ab108335), anti-CHST6 (Abcam, ab154332), anti-N-Cadherin (Abcam, ab76011), anti-Vimentin (Abcam, ab92547), anti-PARP1 (Abcam, ab32138), anti-GAPDH (Proteintech, 60004-1-Ig), anti-Phospho-AKT (Proteintech, 66444-1-Ig), anti-p62 (Proteintech, 66184-1-Ig), Phospho-mTOR (Cell Signaling Technology, #5536), and anti-p-S6 Kinase (Cell Signaling Technology, #9209).