Figure 6.

Failure of VAC formation following AT6V0C knockdown is not due to gross defects in cellular membrane organization prior to infection. (a) Fibroblast cells were transfected with ATP6V0C siRNA or negative control siRNA. At 72 h post-transfection cells were fixed, permeabilised, and stained for early endosomes or trans-Golgi vacuoles. Images are maximum-intensity projections compiled from multiple 0.33 µM slices through the z-axis. (b) Wide field view of fibroblast cells from (a). Image is a single optical slice. (c) Western blot analyses of ATP6V0C siRNA and negative control and transfected fibroblast cells against markers of trans-Golgi vacuoles (TGN46), early endosomes (EEA1). Infected cells were harvested at 72 hpi. (d) Graph shows quantification of representative western shown in C.+ = ATP6V0C siRNA transfected fibroblast, − = negative control siRNA.