Fig. 9. Effect of cholesterol depletion on ER export in the presence of brefeldin A (BFA) and nocodazole. (A) COS7 cells, transfected with VSVG-FP, were shifted to the permissive temperature after overnight incubation at 39.5°C in medium containing LDL-deficient serum and 20 minutes at 4°C when 1 µg/ml nocodazole, 5 µg/ml BFA and 10 mM methyl-β-cyclodextrin were added. Confocal images were captured at 20-second intervals. (B) Analysis of time-dependent average fluorescence intensity (FI) of a single dilated ERES during several cycles of cargo concentration and blink-out in cholesterol-depleted cells. The time zones including an increase in VSVG fluorescence were fitted separately to exponential equations (continuous lines).