Figure 7
From: EV-A71 induced IL-1β production in THP-1 macrophages is dependent on NLRP3, RIG-I, and TLR3
EV-A71 vRNA-triggered IL-1β release is dependent on active caspase-8. (A) Different amounts of EV-A71 viral RNA (0.5, 1 and 2 µg) and 1 µg RD cells RNA were transfected into PMA-primed THP-1 cells for 12 h. The expression levels of EV-A71 5’UTR and IL-1β transcripts were examined by RT-qPCR. (B) Supernatants were collected and the expression of IL-1β was analyzed by ELISA. (C) PMA-primed THP-1 cells were treated with cycloheximide (CHX) (100 μM) for 12 h to suppress the de novo protein synthesis. The untreated and CHX-treated THP-1 macrophages were infected with EV-A71 for 12 h. The expression of EV-A71 3D protein and β-actin was examined by Western blot. (D) The released IL-1β levels were determined by ELISA. (E) THP-1 macrophages were transfected with 1 µg RD cellular RNA or EV-A71 vRNA (0.5, 1, and 2 µg). The expression of pro-caspase-8, cleaved caspase-8, and β-actin was examined by immunoblot. (F) THP-1 macrophages were stimulated by 1 µg RD RNA or EV-A71 vRNA. Then the cells were treated with Z-IETD-FMK for 12 h. The supernatants were harvested and subjected to IL-1β ELISA. (***p < 0.001, Student’s unpaired T-test).
