ESCRT complexes are required for efficient consumption of LDs

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ESCRT complexes are required for efficient consumption of LDs ESCRT complexes are required for efficient consumption of LDs. (A) LD consumption was analyzed in the indicated ESCRT 0 (vps27Δ), I (vps28Δ), II (vps25Δ), III (vps20Δ and snf7Δ) and Vps4p (vps4Δ) mutant and control (wt and tgl3Δ) strains, as described for Fig. 1B (n=3). ESCRT complexes are required for efficient consumption of LDs. (A) LD consumption was analyzed in the indicated ESCRT 0 (vps27Δ), I (vps28Δ), II (vps25Δ), III (vps20Δ and snf7Δ) and Vps4p (vps4Δ) mutant and control (wt and tgl3Δ) strains, as described for Fig. 1B (n=3). (B) Consumption of LDs in WT and indicated mutant cells after dilution from stationary phase into fresh medium was followed by BODIPY493/503 staining and fluorescence microscopy. Scale bar: 5 µm. (C) WT and tgl3Δ mutant cells carrying empty vector, and vps4Δ mutant cells carrying a temperature-sensitive mutant vps4_ts were grown to stationary phase at permissive temperature (26°C) in SC medium lacking leucine. After 30 min pre-incubation at 37°C, cells were diluted into fresh medium containing cerulenin for 5 h at 37°C. LD consumption was analyzed as in Fig. 1B (WT, tgl3Δ n=6; vps4_ts 6 clones, n=6 each). (D) WT and the indicated mutant cells in stationary phase were diluted into fresh medium and their growth was followed by measuring OD (n=3). Data are presented as mean±s.d. ***P<0.001 versus WT (A); *P<0.05 versus WT 37°C and ***P<0.001 versus vps4_ts 26°C (C). Sarah Ouahoud et al. J Cell Sci 2018;131:jcs213876 © 2018. Published by The Company of Biologists Ltd