Known proteins and pathways required for proper mitochondrial morphology and distribution display altered mitochondrial phenotypes as expected. (A) Fluorescent images of known MDM strains were imaged after 24 h of induction. The mitochondria morphology (MitoTracker channel) is severely affected after protein depletion (GFP channel). Scale bar: 5 µm. (B) Schematics of the ergosterol biosynthesis pathway with quantification of the AID strains for the MitoTracker signal (purple no induction, magenta 4 h induction) and GFP (green no induction, gray 4 h of induction). Dashed vertical lines indicate the background fluorescence in the control. Changes are observed irrespective of whether the proteins affected participate in the pre (blue and green) or post (orange) squalene module of the pathway. Image adapted from Jordá and Puig (2020). (C) Fluorescent images of a strain containing the ergosterol sensitive protein, Upc2, C′ tagged with GFP under control conditions, or upon 4 h of treatment with fluconazole or terbinafine, which inhibit ergosterol biosynthesis. Upc2 translocates to the nucleus (green) upon ergosterol depletion. Concomitantly, the MitoTracker signal intensity increases and the mitochondrial morphology changes. (D) Bar graph showing the oxygen consumption rate derived from respiration for a control and selected strains, before and after 7 h of induction with 5-Ph-IAA. Significant changes for each group with the control and for each induced versus their corresponding uninduced group are marked (one-way ANOVA, FDR 0.05). (E) Bar graph showing the oxygen consumption rate not derived from respiration, before and after 7 h of induction with 5-Ph-IAA. Several strains present increased oxygen consumption. Significant changes for each group with the control and for each induced versus their corresponding uninduced group are marked (one-way ANOVA, FDR 0.05).
Figure S4.

Known proteins and pathways required for proper mitochondrial morphology and distribution display altered mitochondrial phenotypes as expected. (A) Fluorescent images of known MDM strains were imaged after 24 h of induction. The mitochondria morphology (MitoTracker channel) is severely affected after protein depletion (GFP channel). Scale bar: 5 µm. (B) Schematics of the ergosterol biosynthesis pathway with quantification of the AID strains for the MitoTracker signal (purple no induction, magenta 4 h induction) and GFP (green no induction, gray 4 h of induction). Dashed vertical lines indicate the background fluorescence in the control. Changes are observed irrespective of whether the proteins affected participate in the pre (blue and green) or post (orange) squalene module of the pathway. Image adapted from Jordá and Puig (2020). (C) Fluorescent images of a strain containing the ergosterol sensitive protein, Upc2, C′ tagged with GFP under control conditions, or upon 4 h of treatment with fluconazole or terbinafine, which inhibit ergosterol biosynthesis. Upc2 translocates to the nucleus (green) upon ergosterol depletion. Concomitantly, the MitoTracker signal intensity increases and the mitochondrial morphology changes. (D) Bar graph showing the oxygen consumption rate derived from respiration for a control and selected strains, before and after 7 h of induction with 5-Ph-IAA. Significant changes for each group with the control and for each induced versus their corresponding uninduced group are marked (one-way ANOVA, FDR 0.05). (E) Bar graph showing the oxygen consumption rate not derived from respiration, before and after 7 h of induction with 5-Ph-IAA. Several strains present increased oxygen consumption. Significant changes for each group with the control and for each induced versus their corresponding uninduced group are marked (one-way ANOVA, FDR 0.05).

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