(A) PJ390 (WT), AN0223 (arn1Δ), PJ380 (tsc2Δ), and AN0224 (tsc2Δarn1Δ) were grown in EMM. (B) PJ390 (WT), AN0223 (arn1Δ), and PJ380 (tsc2Δ) were grown in EMM. Tubulin is shown as a loading control. (C) AN0276 carrying the indicated plasmids was grown in EMM. (D) Cells as in panel C were grown in EMM. (B,D) Proteins were probed with the indicated antibodies. (E) PJ390 (WT) and AN0223 (arn1Δ) were grown in EMM and then treated with 100 µg/ml cycloheximide for 1 hour. (F) PJ390 (WT) and AN0223 (arn1Δ) were grown in EMM and labeled with 50 µM FM4-64 dye on ice for 30 minutes. Five minutes after changing to a fresh medium without the dye at 30°C, the fluorescent signal was observed under a fluorescence microscope. (A,C,E,F) The fluorescence images are shown inverted for clarity. Scale bars: 10 µm.