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. 2013 Sep 16;202(6):875–886. doi: 10.1083/jcb.201305094

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© 2013 Xu et al.

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Figure 5. — Gcs1 is a Drs2 flippase effector. (A and B) Both the GAP domain and ALPS motif are important for Gcs1 function in Snc1 recycling. gcs1Δ cells expressing GFP-Snc1 and an empty vector, wild-type Gcs1, Gcs1 GAP(R54K), or Gcs1 ALPS(L246D) were cultured to mid-log phase, and GFP-Snc1was imaged by fluorescence microscopy. More than 100 cells from three independent isolates of each strain were counted and categorized for GFP-Snc1 localization to the plasma membrane primarily (black bars) or intracellular punctae (gray bars). ***, P < 0.001. Error bars indicate mean + SD. (C) Schematic representation of the Gcs1 constructs used. The region (206–281) containing ALPS was replaced with either a FYVE domain from EEA1 or a PH domain from Osh1. (D) Localization of GFP-Snc1 in drs2Δ cells containing the constructs indicated. Bars, 5 µm.