Skip to main content
. 2019 Nov 6;218(12):3954–3966. doi: 10.1083/jcb.201903006

Figure 1.

Figure 1.

Retromer-dependent cargo recycling of the glutamine transporters ASCT2 and SNAT2. (A) Representative immunoblotting analysis for the indicated proteins in control and VPS35 knockout cells treated with DMSO or bafilomycin A1 (100 nM for 16 h). The quantification represents means ± SEM; n = three independent experiments; one-way ANOVA followed by Dunnett’s multiple comparison. (B) Immunoblot analysis of the total levels of the indicated proteins in control and VPS35 knockout HeLa cells treated with 10 µg/ml cycloheximide (CHX) over the specified time course. Data representative of three independent experiments. (C) Colocalization analysis of endogenous ASCT2 (green) and LAMP1 (red) in control and VPS35 knockout HeLa cells. The graph represents means ± SEM. Pearson’s correlation; n = three independent experiments with >100 cells per condition; Student’s t test (unpaired). (D) IF analysis of endogenous SNAT2 (green), TGN46 (red), and VPS35 (white) in control and VPS35 knockout HeLa cells. (E) Representative IF images of endogenous SNAT2 (green), LAMP1 (red), and VPS35 (white) in control and VPS35 knockout HeLa cells. Graphs in D and E 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 in C–E. *P < 0.05; ****P < 0.0001. KO, knockout.