Fig. 2.

NLRP3 is critical for gp120 LAV-induced inflammasome activation. a–c Western blots of Casp-1-p20 and IL-1β-p17 (a) and the levels of IL-1β (b) and LDH (c, lower panel) in supernatants (Sup) from BV2 cells transfected with control siRNA (si-NC) or NLRP1, NLRP3, AIM2, and NLRC4-specific siRNA and stimulated for 24 h with or without gp120 LAV (0.5 μg/ml). The NLRP1, NLRP3, AIM2, NLRC4 (a) and GSDMD (c, upper panel) in lysates (Lys) of those cells were detected. d–f Western blots of Casp-1-p20 and IL-1β-p17 (d) and the levels of IL-1β (e) and LDH (f, lower panel) in supernatants from BV2 cells transfected with control siRNA or NLRP3, ASC, and Casp-1-specific siRNA and stimulated with or without gp120 LAV (0.5 μg/ml). The NLRP3, ASC, pro-casp-1 (d) and GSDMD (f, upper panel) in lysates of those cells were detected. g ELISA of IL-1β in supernatants of BV2 cells treated with gp120 LAV (0.5 μg/ml) in the presence of various doses of the NLRP3-specific inhibitor (MCC950, 1–25 μM). h GSDMD immunoblot analysis (upper panel) and relative LDH release (lower panel) of BV2 cells treated with gp120 LAV (0.5 μg/ml) in the absence or presence of MCC950 (20 μM). i–k Western blots (i) and densitometric analysis of IκBα (j), NF-κB p65 (k), NLRP3 (l), and pro-IL-1β (m) in BV2 cells treated with the control, gp120 LAV (0.5 μg/ml), LPS + ATP or ATP. The data are displayed as the mean ± SEM (n = 5) from three independent experiments (b, c, e–h, j–m). Western blot results are representative of three (a, c, d, f, h) and five (i) independent experiments. **P < 0.01, ***P < 0.001; NS, no significance