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. 2013 May 27;201(5):673–680. doi: 10.1083/jcb.201212103

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© 2013 Rivera-Molina and Toomre

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Figure 3. — Exocyst rSec8-TagRFP colocalized with endosomal vesicles and remained until full dilation of the fusion pore. (A) Cells treated with Sec8 RNAi and cotransfected with Vamp2-pHluorin (Vamp2-pH) and rSec8-TagRFP were imaged by TIRFM (5 min at 2 Hz). Maximum projection images are shown (Video 3). Vamp2-pHluorin fusion events that colocalize with rSec8-TagRFP are shown as yellow arrowheads and circles; noncolocalizing events are in green and red. White arrows show regions of extensive colocalization in the cell periphery. (B) Galleries of single vesicle fusion of rSec8-TagRFP and different pHluorin (pH) cargos: Vamp2, TfRc, and post-Golgi releasable soluble cargos ssFM4-pH and TfRcFM4-pH. Asterisks and arrowheads show when the fusion pore opens and vesicles fully fuse, respectively. Bar, 2 µm. (C) 120 fusion events were temporally aligned by fusion pore opening and merged (error bars = SEM). (D) Quantification of the percentage of fusion events that contained rSec8-TagRFP for the four cargos. n = 5–8 cells per cargo; 243–428 events/condition; error bars = SD; *, P = 0.001, t test.