Figure 4.

TTP regulates endogenous NLRP3 expression. a, THP-1 were transfected with 40 nm control (siScr_1) or two different TTP siRNA (siTTP_1/2). Starting from 8 h after transfection, cells were stimulated with 12.5 ng/ml of PMA for 0, 4, 8, or 24 h. b, cells were transfected as in a, treated with 12.5 ng/ml of PMA for 4 h as indicated. c, primary human macrophages were transfected with 50 nm control (siScr_1) or TTP siRNA (siTTP_1). Starting from 24 h after transfection, cells were stimulated with 10 ng/ml LPS for 0, 4, or 8 h. d, cells were transfected as in c, treated with 10 ng/ml of LPS for 3 h as indicated. a and c, cells were lysed in SDS sample buffer and NLRP3, Pro-IL-1β, TTP, and β-actin (ACTB) were analyzed by Western blot. Representative blots of at least three independent experiments are shown. b and d, RNA was extracted and gene expression was measured by qPCR. NLRP3, TNF, and IL1B expression was normalized to the geometric mean of ACTB, GAPDH, and RPS13. Mean expression of two independent experiments relative to unstimulated siScr_1-transfected cells ± S.E. is shown.