statistically significant. The reduction in the fraction of Glut1 that colocalises with intracellular markers such as Snx1 or GM130 is consistent with increased localisation of Glut1 to the cell surface, although there does not appear to be a full rescue of the phenotype produced by the D620N mutant. In neither case can the reduced colocalisation of Glut1 with Snx1 or GM130 be due to altered levels of Snx1 or GM130, as any changes observed are not statistically significant (see Fig. 5C,E). We also observed that colocalisation of Glut1 with Lamp1 was reduced after TBC1D5 knockdown (see Fig. S1A), although for this experiment the cells were imaged using a conventional fluorescence microscope and the colocalisation was determined using the Volocity software package. Knockdown of TBC1D5 does appear to enhance Glut1 levels, although the increase was not statistically significant (Fig. S1B).
