Figure 2
From: EV-A71 induced IL-1β production in THP-1 macrophages is dependent on NLRP3, RIG-I, and TLR3
NLRP3 and RIG-I are involved with EV-A71 -triggered IL-1β production in THP-1 macrophage. (A, B) NLRP3 siRNA (siNLRP3) (100 nM) or scrambled siRNA was transfected into PMA-primed THP-1 cells for 48 h. Then the cells were infected with EV-A71 at an M.O.I. of 2 for 12 h. (A) NLRP3, EV-A71 VP0, and β-actin were analyzed by Western blot. (B) The secretion levels of IL-1β were assessed by ELISA. (C, D) PMA-primed THP-1 cells were transfected 100 nM siRNA target RIG-I (siRIG-I) or scrambled siRNA for 48 h, and then infected with 2 M.O.I. EV-A71 for 24 h. (C) The expression levels of RIG-I, EV-A71 VP0, and β-actin were determined by Western blot. (D) Released IL-1β levels were quantified by ELISA. (E, F) Specific ASC siRNA (siASC) and scrambled siRNA were transfected into PMA-primed THP-1 cells for 48 h. Cells were then infected with EV-A71 at an M.O.I. of 2 for 12 h. (E) The expression levels of ASC, EV-A71 VP0, and β-actin were analyzed by Western blot. (F) The supernatants were collected to quantify the released IL-1β protein levels by ELISA. Data are expressed as mean value \(\pm\) SD (***p < 0.001, Student’s unpaired T-test).
