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. Author manuscript; available in PMC: 2014 Jun 11.
Published in final edited form as: Nat Cell Biol. 2013 Apr 7;15(5):461–471. doi: 10.1038/ncb2721

Fig. 3. PDZ interaction motifs at the C-terminus of membrane proteins interact with SNX27 and are required to maintain surface levels.

Fig. 3

(a) The cytoplasmic tails of selected proteins from the surface proteome analysis were fused to GFP and analyzed for binding of endogenous SNX27 and mCherry-tagged SNX27ΔPDZ in immunoprecipitation experiments (uncropped blots shown in Fig. S5) (b) Antibody uptake experiments with antibodies against endogenous CD97 and HA-tagged Glut1. After 1h incubation at 37°, internalized Glut1 and CD97co-localized extensively with endogenous VPS35 or GFP-SNX27 on vesicular structures. (c) Upper panel: Immunofluorescent staining of HA-tagged Glut1 full length and Myc-tagged Glut1 with a truncated PDZ binding motif (Glut1-ΔPDZ), lower panel: Colocalization analysis of HA-Glut1 and HA-Glut1ΔPDZ with the lysosomal marker LAMP1. The graph represents the mean of three independent experiments with 10 images each. (n=3, * indicates p<0.05, unpaired t-test; error bars = s.d., individual data points shown in statistics source data, scale bar = 10μm) (d) Uptake assay with radioactive glucose in SNX27 and VPS35 depleted cells. The graph represents the mean of four independent experiments done in quadruplicates. (n=4,* indicates p<0.05, unpaired t-test; error bars = s.e.m., individual data points shown in statistics source data)