Extended Data Fig. 6: LRP8 boosts TBEV E-mediated attachment to host cells.
a, A549 WT, LRP8-KO and cells overexpressing LRP8 variants (LA1-7, LA1-3, LA1-2, and LA1) were exposed to TBEVSofjin (MOI, 10 IU) at 4 °C. Cells were immunostained for TBEV E, plasma membranes were labelled with wheat germ agglutinin, and cells were imaged by fluorescent confocal microscopy. Images from one experiment representative of >10 randomly captured images of two independent experiments (n = 2) are shown. Scale bar, 50 μm. b (left panel), A549 WT, LRP8-KO and LRP8-overexpressing cells were exposed to rLGTVTBEV-93/783 (MOI, 0.5 IU) at 4 °C and the amounts of bound viral particles were determined by western blotting. GAPDH was included as a loading control. Data from one experiment representative of four independent experiments (n = 4) is shown. Mr, relative molecular weight (K denotes ×1,000). For gel source data, see Supplementary Information Fig. 4. (Right panel), E signal was quantified on a ChemiDoc XRS+ Imaging System using the onboard software. Averages ± s.d. are shown from four experiments, n = 4. WT versus LRP8-KO or LRP8-overexpressing cells, one-way ANOVA with Dunnett’s test: ****, P < 0.0001; ***, P = 0.0003; *, P = 0.0140. c, A549 LRP8-overexpressing cells were exposed to TBEVSofjin (MOI, 10 IU) at 37 °C. Cells were immunostained for TBEV E and the early endosomal marker EEA1 and imaged by fluorescent confocal microscopy. Arrowheads indicate colocalization of TBEV E and EEA1. Images from one experiment representative of >20 randomly captured images of two independent experiments (n = 2) are shown. Scale bar, 50 μm.
