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. 2009 Oct 15;20(20):4278–4288. doi: 10.1091/mbc.E09-03-0243

Figure 5.

Figure 5.

The SNARE Vamp7B localizes to contractile vacuoles and endosomes and is enriched on the contractile vacuoles of AP180-null cells. (A) Cells expressing GFP-Vamp7B (green) were immunostained with the contractile vacuole marker, Rh50 (red), and the endosomal marker, p80 (blue). Contractile vacuoles are indicated by arrows and endosomes are indicated by arrowheads. Scale bar, 10 μm. (B) Vamp7B-containing contractile vacuoles label with clathrin. Wild-type cells expressing GFP-Vamp7B (green) were immunostained with a contractile vacuole marker, Rh50 (blue), and with anti-clathrin light chain (clathrin) (red). Scale bar, 10 μm. (C) Quantification of Vamp7B-labeled contractile vacuoles in wild-type, AP2α−, AP180−, and AP2α/AP180 DKO cells. Error bar, SE, n = 3 independent experiments. In each experiment 32 to 75 contractile vacuoles in each cell line were scored (*p <0.05, **p <0.005). (D) Western blots show equivalent expression of GFP-Vamp7B. In each lane, whole cell lysates (1 × 106 cells) were stained with anti-GFP antibody. Anti–myosin-heavy-chain antibody was used as a loading control.