Figure 2.

Retromer maintains the cell surface abundance and TGN localization of the glutamine transporters ASCT2 and SNAT2, respectively. (A and B) VPS35 knockout HeLa cells were transiently transfected with GFP-VPS35 or GFP alone. After 48 h of incubation, cells were fixed and immunostained with antibodies to endogenous ASCT2 (green) and LAMP1 (red; A) or SNAT2 (green) and LAMP1 (red; B). (C) Colocalization analysis of endogenous SNAT2 (green) and TGN46 (red) in VPS35 knockout HeLa cells transiently transfected with GFP-VPS35 or GFP alone. White asterisks indicate transfected cells. Graphs represent means ± SEM. Pearson’s correlation of three independent experiments with >150 cells per condition; Student’s t test (unpaired). Scale bars, 20 µm and 10 µm in magnified merge panels. ***P < 0.001; ****P < 0.0001.