Fig. 4: APIP stimulates the priming of NLRP3 inflammasome activation.

a–f qRT-PCR analysis of LPS-induced inflammation. J774A.1 cells were transfected with negative control siRNA (NC) or siApip (100 nM, 48 h) and left untreated (NT) or treated with LPS (500 ng/ml, 3 h) (a–d). Control and Apip cKO BMDMs were treated with LPS (200 ng/ml, 3 h) (e, f). Total RNAs were analyzed by qRT-PCR. Data represent mean ± SD (n = 3 independent cultures). g–k Western blot analysis of LPS-induced inflammation. J774A.1 cells were transfected with NC or siApip (100 nM, 48 h) and treated with LPS (500 ng/ml, 3 h) (g, h). RAW264.7 cells were transfected with Mock or APIP for 24 h and treated with LPS (1 μg/ml) alone or with Takinib (10 μM) for 3 h (i–k). Cell lysates were analyzed by western blotting (g, i), and blot signals quantified by densitometry (h, j, k). Data represent mean ± SD (n = 4 independent cultures; h n = 5 for pro-IL-1β and n = 3 for p-IκBα). Statistical tests: two-way ANOVA with Tukey’s multiple comparison test (a–d, j), unpaired two-tailed Student’s t-test (e, f), multiple unpaired t-test with Holm–Sidak’s method (h), or one-way ANOVA with Tukey’s multiple comparison test (k). ns, non-significant. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001, ^****p < 0.0001. Exact p values are in the Source Data. Source data are provided as a Source Data file. arb. units, arbitrary units.