Fig. 6.

ARF6 inactivation is sufficient to inhibit nutrient transporter recycling. (A) HeLa cells expressing HA-ARF6 or HA-ARF6-T27N stained for CD98. (B) Quantification of intracellular CD98 intensity in A. (C) Surface CD98 quantification and ARF6-GTP levels in FL5.12 cells treated with FTY720 (5 µM) or SecinH3 (30 µM) for 3 h. (D) HeLa cells treated with SecinH3 (30 µM) or NAV2729 (12.5 µM) for 16 h and stained for CD98. (E) HeLa cells treated with SecinH3 (30 µM) or NAV2729 (12.5 µM) for 1 or 6 h and stained for MICAL-L1. (F) Quantification of E. (G) Cytoplasmic intensity quantification of CD98 and GLUT1 in HeLa cells treated with vehicle, 893 (10 µM), or C₂-ceramide (50 µM) for 1 h, washed with PBS, then returned to 893 (10 µM) or placed in vehicle or SecinH3 (30 µM) for 2 h. Means±s.e.m. shown for n≥3 in all panels. Using a two-tailed t-test (B) or an ordinary one-way ANOVA (C,F,G) to compare treated samples with controls: n.s., not significant; *P≤0.05; **P≤0.01; ***P≤0.001. Dunnett's test was used in C,F and Tukey's test was used in G to correct for multiple comparisons. Scale bars: 10 µm. See also Figs S6 and S7.