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. 2003 Oct 1;4(12):1182–1189. doi: 10.1038/sj.embor.7400018

Figure 6.

Figure 6

Sorting of VAMP4 in AtT20 cells requires both AP-1 and PACS-1 binding sites. The localization of transfected Flag-tagged (A) wt VAMP4, (B) VAMP4 LL/VV, (C) VAMP4 S30A or (D) VAMP4 LL/VV S30A was compared to endogenous ACTH in AtT20 cells. The left panels show the Flag tag (green, M2 ab), the centre panels show ACTH (red, POMC ab) and the right panels show the MERGE of the two images. The wt, LL/VV and S30A proteins remain predominantly in a perinuclear region, and are not found in the tips (arrowheads in (A–C)) with ACTH. Mutation of both the dileucine motif and the PACS-1 binding motif results in mislocalization of VAMP4 to ACTH-positive MSGs found in the tips of the cells (arrowhead in (D)). The inset in (C) MERGE shows double labelling of VAMP4 and ACTH in another tip.