To load lysosomes, HeLa cells were preincubated with dextran-488 o/n, then chased in dextran-free media for 3 h before infection with Red-

<p><b>Copyright information:</b></p><p>Taken from " Trafficking is Defined by Continuous Dynamic Interactions with the Endolysosomal System"</p><p></p><p>Traffic (Copenhagen, Denmark) 2007;8(3):212-225.</p><p>Published online 15 Jan 2007</p><p>PMCID:PMC2063589.</p><p>© 2007 The Authors Journal compilation © 2007 Blackwell Publishing Ltd</p> To follow endocytic access to the SCV dextran-488 was added various times after infection. Cells were then observed by confocal microscopy. Images of dextran-488 and Red- were overlaid and the number of SCVs containing dextran-488 were counted. A) Time course of SCV interaction with terminal lysosomes (filled bars) and incoming endocytic traffic (open bars). B) To assess the effect of microtubule depolymerization or V-ATPase inhibition on marker acquisition 20 μM nocodazole or 0.5 μM bafilomycin was added, respectively, 1 h before infection and maintained throughout the experiment. Live-cell imaging was initiated at 2 h p.i. and SCVs containing dextran-488 were counted. Values are the mean ± SD of three independent experiments ( ≥ 60 in each experiment). Asterisk indicates significant difference from control-treated cells (p < 0.05, , Tukey’s test)