Fig. 1.

Natural and synthetic sphingolipids trap plasma membrane nutrient transporters along with recycling cargo. (A) FL5.12 cells were treated with vehicle, C₂-ceramide (50 µM), bacterial sphingomyelinase (bSMase; 100 mU/ml), FTY720 (5 µM) or SH-BC-893 (893, 5 µM) for 3 h then stained for surface CD98. (B) HeLa cells were treated as in A for 12 h then stained for CD98. All images were collected with the same microscope and processed using the same settings. (C-F) HeLa cells were treated with vehicle, C₂-ceramide (50 µM), or 893 (10 µM) for 12 h, then stained for CD98 and EEA1 (C), CD98 and TGN46 (D), GLUT1 and transferrin receptor (TfR) (E), or CD98 and LAMP1 (F). n≥3 for all panels. Data in A are means±s.e.m. One-way ANOVA was used to compare treated samples with control, ***P≤0.001. Dunnett's test was used to correct for multiple comparisons. Scale bars: 10 µm. See also Figs S1-S5.