Figure 4

Effects of A1AT-0 and A1AT-LA on LPS-induced NLRP3 expression. Human neutrophils (5 × 10⁶) were incubated for 5 h in either medium alone or medium containing LPS (20 ng/mL) in the absence or presence of A1AT-0 (1 mg/mL) or A1AT-LA (1 mg/mL). mRNA expression of the NLRP3 gene (A) was analyzed by real-time qPCR as described in Materials and Methods. GAPDH and GUSB were used as housekeeping genes. Box plots represent data from 4 individual donors; n = number of replicates for each experiment. P value indicates significant differences compared with the values seen in LPS-activated cells. Total cell lysates were analyzed for NLRP3 protein levels by western blotting (B). For loading control, blots were reprobed with antibodies to β-actin. Each blot is representative of 3 independent donors with similar results.