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. Author manuscript; available in PMC: 2017 Nov 1.
Published in final edited form as: Mol Microbiol. 2016 Aug 25;102(3):488–505. doi: 10.1111/mmi.13474

Figure 7.

Figure 7

(A) The ΔgcsA strain is unable to synthesize GlcCer. The ΔsdeA, ΔsmtA, ΔsmtB, ΔgcsA and parental conidia were grown in YUU medium at 37 °C for 3-4 days. The total lipids were obtained according to Bligh & Dyer and submitted to TLC using chloroform: methanol: water 65:25:4 as the solvent mixture. Bands corresponding to distinct lipid classes were visualized after iodine staining, and 25 μg of soy GlcCer was used as the migration standard of the molecule (arrow). (B, C) The relative abundance of the intermediates involved in GlcCer synthesis after growth in YUU media (B) or under niiA-inducing/repressing conditions (C). (B) The relative abundance of α-OH-Δ4-ceramide (hatched light grey bars), α-OH-Δ4-Δ8-ceramide (hatched dark grey bars), α-OH-Δ4-Δ8-9,methyl-ceramide (hatched black bars) and their glycosylated products (α-OH-Δ4-GlcCer, filled light grey bars; α-OH-Δ4-Δ8-GlcCer, filled dark grey bars; and α-OH-Δ4-Δ8-9,methyl-GlcCer, filled black bars, respectively) in the ΔsdeA, ΔsmtA, ΔsmtB, ΔgcsA and wild-type lipid extracts. All of the values represent the mean ± SEM of two independent experiments, as analyzed by a one-way ANOVA. αp < 0.001, ΔsdeA versus WT; βp < 0.001, ΔgcsA versus WT; χp < 0.001, ΔsdeA versus WT; δp < 0.01, ΔsmtA versus WT; εp < 0.05, ΔsmtB versus WT; φp < 0.05, ΔsmtB versus WT). (C) Relative abundance of GlcCer synthesis intermediates in the wild-type, ΔsmtA and ΔsmtA niiA::smtB strains grown in MMM medium supplemented with 10 mM sodium nitrate (black, hatched light grey and hatched dark grey bars, respectively) or 50 mM ammonium tartrate (filled black, filled light grey and filled dark grey bars, respectively) for niiA-induction/repression. The values represent the means ± SEM of three independent experiments, as analyzed by a one-way ANOVA. αp < 0.001, ΔsmtA niiA::smtB + ammonium tartrate versus wild-type + ammonium tartrate, βp < 0.001, ΔsmtA niiA::smtB + ammonium tartrate versus ΔsmtA + ammonium tartrate, χp < 0.01, ΔsmtA niiA::smtB + ammonium tartrate versus ΔsmtA niiA::smtB + sodium nitrate; δp < 0.001, ΔsmtA niiA::smtB + ammonium tartrate versus wild-type + ammonium tartrate, εp < 0.001, ΔsmtA niiA::smtB + ammonium tartrate versus ΔsmtA + ammonium tartrate, φp < 0.001, ΔsmtA niiA::smtB + ammonium tartrate versus ΔsmtA niiA::smtB + sodium nitrate.