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. 2022 May 24;221(4):iyac086. doi: 10.1093/genetics/iyac086

Fig. 6.

Fig. 6.

Cellular acyl-CoA chain length and FAS activity partially modulate eaf1Δ cell sensitivity to Sor A. a) FAS activity is increased in eaf1Δ cells compared to wt cells, but Cerulenin inhibits FAS activity in both cells. Wild-type (YKB 1079) and eaf1Δ (YKB 3333) were grown to early-log phase at 30°C in YPD. Cells were then again diluted to an OD600 of 0.1 in YPD with or without 2.5 µM Cerulenin, and then grown for 2 h at 30°C. FAS activity assay was performed on the whole-cell extracts and was normalized to the relative protein abundance in the sample. Error bars denote the standard error of the mean (SEM). n = 3, *P < 0.05 determined using a 2-way ANOVA test with a Tukey’s multiple comparisons test. b) Cerulenin partially suppresses sensitivity of eaf1Δ cells to Sor A. eaf1Δ (YKB 4448) cells were grown to early-log phase at 30°C in YPD. Cells were then diluted to an OD600 of 0.1 in YPD with or without 0.075 µM Sor A and/or 2.5 µM Cerulenin, and an automated growth curve analysis was performed and used to calculate growth rate. Error bars denote the standard error of the mean (SEM). n = 3, *P < 0.05 determined using a 2-way ANOVA test with a Sidak’s multiple comparisons test. c) Palmitic acid (C16) increases sensitivity of eaf1Δ cells to Sor A. eaf1Δ (YKB 4448) cells were grown to early-log phase before being diluted to an OD600 of 0.1 in YPD containing 0.1% Brij 58 with or without 0.075 µM Sor A and/or 0.8 mM palmitic acid (C16) or 0.8 mM stearic acid (C18), automated growth curve analysis was performed at 30°C for 48 h and growth rate was then calculated from 3 biological replicates. Error bars denote the standard error of the mean (SEM). n = 3, *P < 0.05 determined using a 2-way ANOVA test with a Sidak’s multiple comparisons test. d) Palmitic acid induces the formation of thick rod-like structures of Acc1-GFP in eaf1Δ cells. eaf1Δ (YKB 4448) cells expressing endogenously tagged Acc1-GFP were grown to early-log phase before being diluted to an OD600 of 0.1 in YPD containing 0.1% Brij 58 with or without 0.8 mM myristic acid (C14), 0.8 mM palmitic acid (C16), or 0.8 mM stearic acid (C18) and grown for 2 h at 30°C prior to imaging. Representative brightfield and fluorescent images are shown. Scale bar: 4 µm.