Z-stacks (150 cells for each condition and time point) comprising the total height of the cells were acquired (X sections at 0.5 mm intervals) to measure EGFP and anti-VSVG fluorescence as described for the Golgi analysis

Before immunostaining, blots ended up stained for overall protein with Ponceau S (Sigma) they had been then incubated with antibodies diluted in TBS+5% milk+.1% Tween. Peroxidase-conjugated secondary antibodies ended up…