FIG 9.

NAP1L1 controls the TLR3 pathway. (A) Depletion of NAP1L1 affects TRIF-mediated activation of IFN-β. Huh7-Lunet cells were transduced with LV for shNAP1L1 or shCTRL and subsequently transfected with expression vectors for TRIF together with the reporter IFN-β-Luc and the Renilla control. Luciferase activity was measured in triplicate independent experiments, normalized for Renilla, and represented as fold change over mock treatment ± SD. (B) Depletion of NAP1L1 by LV shRNA treatment. Huh7-Lunet cells were transduced with LV for shNAP1L1 or shCTRL and then with LV expressing TLR3 or EGFP as a control. Fifty micrograms of poly(I·C) was added to the medium and left for 24 h. IFN-β mRNA levels for NAP1L1 were measured by qRT-PCR, normalized for β-actin, and plotted against those for mock treatment. Averages for 3 independent replicates are shown with standard deviations. (C) Reconstitution of the TLR3 pathway. Cells were processed and TLR3 mRNA quantified as for panel B. (D) Depletion of NAP1L1 affects TLR3 signaling. Cells were processed and IFIT1 mRNA quantified as for panel B.