Table II.

Quantitation of HRP Label in Pericentriolar ARF6-positive Vesicles Relative to Other Small Vesicles (<200-nm Diameter)

Cells transfected with		Small vesicles (<200 nm)				ARF6-positive vesicles
		Minutes post uptake
		5		60		5		60
ARF6		34		173		0		12
ARF6(Q67L)		21		112		0		0
ARF6(T27N)		59		189		0		28
Vector		89		342		−		−

CHO cells on monolayers expressing ARF6 wild-type or mutant proteins were incubated with HRP as described in Materials and Methods. Monolalyers were quickly rinsed, fixed, and processed for cryoimmunogold labeling and doubled-labeled with antibodies directed against ARF6 followed by protein A–10-nm gold and against HRP followed by protein A–5-nm gold as described in Materials and Methods. The data represents an average of 5-nm gold particle count obtained from 25 cells for each condition.