Anti human aim2

Cells were harvested and lysed in RIPA buffer (Cell Signaling Technology, Danvers, MA, USA) containing protease inhibitor cocktail (Sigma-Aldrich). Proteins samples were separated using 10–15% SDS-PAGE and then transferred to membranes (Millipore, Burlington, MA, USA), which were then probed with primary and secondary antibodies. Membranes were developed using a chemiluminescence solution (GE Healthcare, Chicago, IL, USA) in a LAS-4000 Lumino-imaging unit (Fujifilm, Tokyo, Japan). Immunoblot band intensities were quantified using NIH ImageJ software (Version 1.53t, Fujifilm), and results were presented as intensity ratios versus β-actin. The antibodies used were as follows; anti-β-actin (Santa Cruz Biotechnology, Dallas, TX, USA), anti-human iNOS (Santa Cruz Biotechnology), anti-human TLR2 (Santa Cruz Biotechnology), anti-human TLR4 (Santa Cruz Biotechnology), NF-κB (Cell Signaling Technology), MAPK Family antibody sampler kit (Cell Signaling Technology), anti-human AIM2 (Cell Signaling Technology), anti-human ASC (Cell Signaling Technology), pro-Caspase-1 (Cell Signaling Technology), anti-human IL-1β (R&D Systems, Minneapolis, MN, USA), Beclin1 (Santa Cruz Biotechnology), ATG5 (Santa Cruz Biotechnology), and LC3 (Cell Signaling Technology).

Phorbol 12-mystristate 13-acetate (PMA), oxidised ATP (oxATP), KCl, and trichloroacetate (TCA) were purchased from Sigma (St. Louis, MO, USA). CA-074 methyl ester (CA-074Me) was obtained from Calbiochem (San Diego, CA, USA). Z-VAD-FMK(pan-caspase inhibitor), Z-WEHD-FMK(caspase-1 inhibitor), anti-human IL-1β, and anti-human NLRP3 were supplied by R&D Systems (Minneapolis, MN). Anti-human ASC, anti-human AIM2, and anti-human caspase-1 antibodies were acquired from Cell Signaling Technology (Beverly, MA, USA). The anti-human NLRC4 and anti-β-actin antibodies were procured from Santa Cruz (Santa Cruz, CA, USA).

Cells were harvested and lysed in RIPA buffer (Cell Signaling Technology, Danvers, MA, USA) containing protease inhibitor cocktail (Sigma-Aldrich). Protein samples were separated using 10–15% SDS-PAGE and then transferred to membranes (MilliporeSigma, Burlington, MA, USA), which were then probed with primary and secondary antibodies. Membranes were developed using a chemiluminescence solution (GE Healthcare, Chicago, IL, USA) in a LAS-4000 Lumino-imaging unit (Fujifilm, Tokyo, Japan). Immunoblot band intensities were quantified using NIH ImageJ 1.54f software (Fujifilm), and the results are presented as intensity ratios versus β-actin. The antibodies used were as follows: anti-β-actin (Santa Cruz Biotechnology, Dallas, TX, USA), anti-human AIM2 (Cell Signaling Technology), anti-human ASC (Cell Signaling Technology), pro-Caspase-1 (Cell Signaling Technology), anti-human IL-1β (R&D Systems, Minneapolis, MN, USA), and NLRP3 (NOVUS biologicals, Centennial, CO, USA)