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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. 4. Suppression of retromer SNX-BAR proteins does not phenocopy SNX27 and VPS35 loss of function but appears to cause trafficking delays.

Fig. 4

(a) HeLa cells were transfected with the indicated siRNAs and surface abundance of Glut1, ATP7A and Steap3 was determined by quantitative western blotting (graph represents the mean of 3 (ATP7A), 6 (GLUT1) and 5 (STEAP3) independent experiments, * indicates p<0.05, unpaired t-test; error bars = s.e.m., individual data points shown in statistics source data, uncropped blots shown in Fig. S5) (b) Immunofluorescent colocalization analysis of endogenous Glut1 and LAMP1 in SNX27-retromer and SNX-BAR depleted cells. The graph represents the mean of 30 images acquired in three independent experiments (n=3, * indicates p<0.05, unpaired t-test; error bars = s.d., Scale bar = 10μm, individual data points shown in statistics source data) (c) Colocalization analysis of endogenous Glut1 and the early endosome marker EEA1 in SNX-BAR depleted cells. 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., Scale bar = 10μm, individual data points shown in statistics source data) (d) Mean of four degradation assays of Glut1 and ATP7A in SNX-BAR depleted cells (for assay details see Fig. 2D or methods, n=4, * indicates p<0.05, unpaired t-test; error bars = s.e.m., individual data points shown in statistics source data, uncropped blots shown in Fig. S5)