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. 2016 Aug 1;214(3):293–308. doi: 10.1083/jcb.201605090

Figure 6.

Figure 6.

A cohort of VARP localizes to melanosomes and accompanies GFP-VAMP7 on departing tubules. (a–i) WT melan-Ink4a melanocytes transiently transfected with VARP-GFP and mCh-VAMP7 (a–c), GFP-VAMP7 and VARP-HA (d–f), or VARP-GFP and mCh-STX13 (g–i) were fixed 48 h later, labeled with anti-HA (d–f), and analyzed by deconvolution immuno-FM. BF images are pseudocolored blue and shown in merge and insets (boxes magnified five times). Arrowheads show VARP puncta adjacent to melanosomes (a–f) or to mCh-STX13–labeled endosomes (g–i). Bars: (main), 10 µm; (insets), 2 μm. (j–u) WT melan-Ink4a melanocytes transiently transfected with VARP-GFP and either mCh-VAMP7 (j-l; see Fig. S3 a), mRFP-OCA2 (m–o; see Fig. S3 b) or mCh-STX13 (p–u; see Fig. S3, c and d) were analyzed 48 h later by spinning-disk confocal microscopy at ∼1 fps. Elapsed time (in seconds) is indicated at lower right. (j–l) A VARP-GFP– and mCh-VAMP7–labeled tubule (arrow) extends from a melanosome (arrowhead). (m–o) A VARP-GFP–labeled vesicle (arrow) exits from an mRFP-OCA2–labeled melanosome (arrowhead). (p–r) A VARP-GFP tubule (arrow) departs from a mCh-STX13–labeled endosome (arrowhead). (s–u) A mCh-STX13 tubule (arrow) exits from a mCh-STX13/VARP-GFP double-labeled endosome (arrowhead). Bars, 2 µm.