Fig. 2.
The Lrp1-binding chaperone RAP can inhibit OROV infection of Vero E6 cells and undifferentiated SH-SY5Y cells. (A) RAP is a 39-kDa ER-resident protein consisting of 3 domains (D1-3) that chaperones LDLR family proteins, including LRP1. Recombinant mRAPD3 and mutant mRAPD3 (K256A and K270E) were expressed and purified from BL21 (DE3) cells using an N-terminal His-tag. mRAPD3 or mutant mRAPD3 was added to (B) Vero E6 nonhuman primate cells or (C) SH-SY5Y human neuroblastoma cells 1 h before infection with MOI 0.1 of RVFV, OROV, or ZIKV. Samples were harvested at 24 hpi (for RVFV and OROV) or 48 hpi (for ZIKV), and infectious virus was measured by plaque assay or qRT-PCR. Statistical significance was determined using two-way ANOVA on log-transformed data. Experiments were repeated three times. **P < 0.01; ****P < 0.0001.