Figure 5.

Loss of VAC due to ATP6V0C1 knockdown is quantifiable and significant. (a) Fibroblast cells were transfected with ATP6V0C siRNA or negative control siRNA and at 72 h post-transfection were infected with AD169. At 144 hpi cells were fixed, permeabilised and stained for early endosomes or trans-Golgi vacuoles, (EEA1:green), viral tegument protein (pp28:red) and nuclei (DAPI:blue). Images represent single slices through the Z-axis. (b) Representative scatter-plot showing average pixel signal intensity in red (pp28) and green (EEA1) channels from multi-cell images (n = 16 for ATP6V0C and n = 9 for negative siRNA). Individual images in the Z-field were analysed using Fiji image analysis software. (c) Pearson's R-value for colocalization of TGN46 or EEA1 and pp28 in ATP6V0C or negative control siRNA transfected fibroblast cells (n = 20). *p-value < 0.05; **p-value < 0.01.