FIG. 9.
CA074R GP17K intermediates require cysteine cathepsin activity for entry. Vero cells were pretreated with 1% DMSO or 300 μM E64 for 4 h, challenged with the THL-derived WT or mutant rVSV-GP17K at an MOI of 0.001 FFU/cell, and then overlaid with agarose. Viral titers (FFU/ml) were determined at 18 hpi. Averages ± standard deviations from one representative experiment are shown. An aliquot of each cleaved virus was subjected to SDS-PAGE, and GP1 was detected by Western blotting to confirm GP cleavage. The higher mobility of GP1 fragments from N40K and T42A reflects their loss of N-glycan at N40. The position of the ∼17-kDa GP1 cleavage fragment is indicated on the left.