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. 2015 Jan 20;112(5):1434–1439. doi: 10.1073/pnas.1409476112

Fig. 5.

Fig. 5.

FAS degradation is important for cell survival under nitrogen starvation. (A) fas1∆ (TOS029) and fas2∆ (TOS030) S. cerevisiae cells were grown to midlog phase and incubated in YPD, SD-N, YPD + 0.1 mM palmitic/stearic/myristic acids, or YPD + 200 nM rapamycin. Cell viability was determined after 16 h using phloxine B. (Scale bar, 10 µm.) (B) Quantification of A. More than 300 cells were counted for each experiment. Error bars represent the SDs of three independent experiments. ***P < 0.001 (Student’s t test) (Right). (C) WT (BY4741) cells were grown to midlog phase and incubated with DMSO, 50 μM cerulenin (cer) or 50 μM cerulenin + 0.1 mM palmitic/stearic/myristic acids (cer + FA). Cell viability was determined after 16 h using phloxine B. More than 300 cells were counted for each experiment. Error bars represent the SDs of three independent experiments. n.s., not significant; ***P < 0.001 (Student’s t test). (D) WT (BY4741) and atg1∆ (TOS001) cells were grown to midlog phase and shifted to SD-N medium for 16 h. FAS activity was determined at the indicated time points as described in Materials and Methods. Error bars represent the SDs of three independent experiments. n.s., not significant; ***P < 0.001 (Student’s t test). (E) WT (BY4741) and DaMP-fas1 (TOS031) cells were grown to midlog phase. Cells were treated with either DMSO or 50 μM cerulenin for 30 min. FAS activity was determined as described in Materials and Methods. Error bars represent the SDs of three independent experiments. ***P < 0.001 (Student’s t test). (F) WT (BY4741), atg7∆ (TOS005), DaMP-fas1 (TOS031) and DaMP-fas1 atg7∆ (TOS032) cells were grown to midlog phase and shifted to SD-N medium. Cell viability was determined at the indicated time points using phloxine B. More than 300 cells were counted for each experiment. Error bars represent the SDs of three independent experiments. n.s., not significant; **P < 0.01 (Student’s t test). (G) WT (BY4741) and atg7∆ (TOS005) strains were grown to midlog phase and shifted to SD-N medium in the presence of DMSO or 50 μM cerulenin. Cell viability was determined using phloxine B at the indicated time periods. Error bars represent the SDs of three independent experiments. n.s., not significant; *P < 0.05, **P < 0.01, ***P < 0.001 (Student’s t test).