Figure 9.

Transfection of ATP6V1A or its truncate trans-complements decreased ERA uncoating in ATP6V1A depleted cells. HEK293T cells were transfected with ATP6V1A siRNA or scrambled siRNA. At 48 h post transfection of siRNA, cells were transfected with the indicated plasmids for 24 h and infected with ERA at an MOI of 100 for 1 h on ice. Cells were then treated with PBS (pH 5) for 15 min, washed, and incubated at 37 °C for 2 h in the presence of cycloheximide. RABV M protein (green) and cell nuclei (blue) were stained and observed for M protein aggregation by confocal microscopy. Yellow solid arrows indicate high-intensity punctate staining of M proteins (A). Quantitative analysis of RABV uncoating in virus-infected cells. On the basis of the confocal microscopy in A, the infected cells at 3 h post infection was categorized into two types, containing more than 10 high-intensity punctate staining spots in each cell, or containing less than 10 spots in each cell. The results shown were calculated from 130 cells under a confocal microscope with a 20× objective lens (B).