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Recombinant Proteins> | NLRP3 in SMG cell lysates (500-1,000 µg protein/ml) was immunoprecipitated with protein A MagBeads (GenScript, Piscataway, NJ) pre-coated with mouse anti-mouse NLRP3 antibody (1:200 dilution; Cryo-2, Cat. No. AG-20B-0014-C100, Adipogen, San Diego, CA) by incubation for 1 h at room temperature. Immunoprecipitated proteins were solubilized using 40 µl of 1X SDS sample buffer (62.5 mM Tris-HCl at pH 6.8, 2% (w/v) SDS, 10% (v/v) glycerol, 50 mM DL-DTT and 0.01% (w/v) bromophenol blue) followed by heating at 95°C for 5 min. The samples were then subjected to SDS-PAGE on a 12% (w/v) gel (GenScript, Piscataway, NJ) and then transferred to nitrocellulose membranes for Western blot analysis. | Get A Quote |
Salivary gland inflammation is a hallmark of Sjögren’s syndrome (SS), a common autoimmune disease characterized by lymphocytic infiltration of the salivary gland and loss of saliva secretion, predominantly in women. The P2X7 receptor (P2X7R) is an ATP-gated non-selective cation channel that induces inflammatory responses in cells and tissues, including salivary gland epithelium. In immune cells, P2X7R activation induces the production of proinflammatory cytokines, including IL-1β and IL-18, by inducing the oligomerization of the multiprotein complex NLRP3-type inflammasome. Here, our results show that in primary mouse submandibular gland (SMG) epithelial cells, P2X7R activation also induces the assembly of ... More