FIG 6 .

Reduction in cellular membrane cholesterol delays, but does not block, viral internalization. (A) Schematic of the virus internalization assay. (B) U2OS cells were treated with the S1P inhibitor (25 µM) or the 1% DMSO vehicle for 24 h and exposed to VSV(mNG-P)-ANDV GP particles at 4°C. Following internalization of virus at 37°C for the indicated times (in minutes), remaining surface-bound virus particles were removed by a brief trypsin treatment. Cells were harvested and fixed with 4% PFA, and internalized virus was measured by microscopy (B) and flow cytometry (C and D) (n = 4; two independent experiments).