Fig. 3.
Lrp1 KO reduces VSV-OROV infection in BV2 cells and VSV-OROV binds to Lrp1 CLIV. BV2 WT and BV2 Lrp1 KO cells were infected with MOI 1 of VSV or MOI of 5 of VSV-OROV. Samples were collected at 6 and 8 hpi to be processed by (A) flow cytometry or (B) imaging by fluorescent microscopy (20×). Scale bars, 50 μm. (C) LRP1 consists of a 515-kDa extracellular alpha chain (blue/tan) and an 85-kDa intracellular beta chain (not shown) connected by a transmembrane domain (gray). The alpha chain is further divided into four complement-type repeat clusters (CLI-IV; blue), and epidermal growth factor (EGF)-like and YWTD domains (tan). Recombinant Fc-fused LRP1 CLII and CLIV were expressed and purified from Expi293 cells for the experiments presented here. (D) AHC sensors coated with either Fc or Lrp1-CLIV-Fc and incubated with VSV-OROV particles. Sensograms show the over-time association and dissociation of virus particles to coated sensors. Significance was determined using an unpaired t test. Experiments were repeated two times. **P < 0.01; ****P < 0.0001.