Fig. 8: VSV-S triggers activation of phospholipid scramblase and the translocation of phosphatidylserines.

a Vero cells were mock-infected or infected with the indicated viruses for 30 min in the absence or presence of CIMSS or staurosporine (10 μM each) or murine anti-ACE2 (anti-ACE), anti-Spike (anti-S) or an isotype control IgG (10 μg/ml of each immunoglobulin). The cells were lysed and incubated with rabbit anti-PLSCR1 antibody and immune complexes precipitated with protein A-agarose and analyzed by western blotting with a mouse anti-phosphotyrosine (PY20) or mouse anti-PLSCR mAb. The blot is representative of results obtained in two independent experiments. b Vero cells were stained for plasma membranes (green) with wheat germ agglutinin conjugated with Alexa488 and then mock-infected or infected with the indicated viruses in the presence of 0.1% DMSO or 10 μM CIMSS and after 30 min, 1 h, or 4 h, fixed and stained with an antibody to phosphatidylserines (PtdS) (red); nuclei were stained with DAPI. Images were obtained with Leica SP8 (objective 63 × 1.4, bar = 10 μm). c Vero cells were mock-infected or infected with the indicated VSV pseudotyped viruses in the absence or presence of 10 µM CIMSS. After 15, 30, or 60 min, the cell surface proteins were biotinylated and precipitated with streptavidin magnetic beads and analyzed by immunoblotting with Abs to pAkt^T308 and total extracellular Akt. Results are representative of two independent experiments.