Western Blotting Analysis of Inflammatory Proteins

Western blotting was performed as we described previously [13 (link)]. The primary antibodies were used as follows: rabbit anti-NALP3 (1 : 1000; Protein Tech Group, Chicago, IL), rabbit anti-ASC (1 : 100; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-caspase-1 (1 : 100; Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse anti-desmin (1 : 1000; Protein Tech Group, Chicago, IL), rabbit anti-synaptopodin (1 : 1000; Protein Tech Group, Chicago, IL), mouse anti-TXNIP (1 : 1000; MBL, International Co, Woburn, MA, USA), goat anti-gp91^phox, and mouse anti-β-actin (1 : 10000; Santa Cruz Biotechnology Santa Cruz, CA, USA). The membrane was incubated with primary antibodies overnight at 4°C, followed by incubation with horseradish peroxidase-labeled IgG (1 : 10000) at room temperature for 1 hour. The immunoreactive bands were detected by chemiluminescence methods. Densitometric analysis of the images was performed by using Image J software (NIH, Bethesda, MD, USA).