also demonstrated an increase in endosomal VPS26 after TBC1D5 knockdown, as these cells contain endogenous Rab7a in addition to the stably expressed GFP-tagged version. Knockdown of Rab7a reduced the VPS26 fluorescence intensity, except for experiments where Rab7a was expressed as a GFP-tagged protein: as the Rab7a in this construct is of murine origin, it is resistant to the Rab7a siRNA we used. The gain in total fluorescence for VPS26 was again not due to increased spot number (see Fig. 2D).
