Figure S2.
Ybr063c (Cnm1) does not affect the extent of ERMES-mediated contacts. (A) ERMES components Mmm1 or Mdm34 were C terminally tagged with GFP on the background of a control strain or strains overexpressing YBR063C (TEF2pr-YBR063C) or deleted for it (Δybr063c). Overexpression of Ybr063c resulted in clustering of ERMES signal to the nuclear ER area but did not change the number or intensity of ERMES puncta. Deleting ybr063c had no effect on these proteins. Scale bar, 5 µm. (B) Ybr063c can be found in distinct areas from ERMES subunits. Overexpressed Ybr063c was N terminally tagged with mCherry on the background of Mdm34 or Mmm1 C terminally tagged with GFP. The yellow arrows represent areas of proximity between the Ybr063c signal and the ERMES proteins, while the white arrows represent areas of Ybr063c signal that does not colocalize with ERMES. Scale bar, 5 µm. (C) A spot dilution assay of strains expressing ybc063c under a GAL promoter in control strains and strains that harbor deletions in mdm34 or vam6. Repressed expression of ybr063c when controlled under the GALpr and grown in glucose caused a complete rescue of the growth defect of Δvam6 in glucose. In contrast, repressing ybr063c on the background of Δmdm34 aggravated the severe growth phenotype of this strain. All strains were grown on both synthetic media with glucose (no expression of Ybr063c) or galactose (Ybr063c is expressed) as a control. (D) 100 representative samples of either the nucleus (on the left) or mitochondria (on the right) that were considered in the quantification analysis of Fig. 3 D. The nuclei were marked by Nsg1-GFP, while the mitochondria were dyed using MitoTracker Orange. (E) Representation of the overlap analysis between the nucleus and mitochondria by artificial intelligence algorithms (ScanR Olympus soft imaging solutions, version 3.2). Mitochondria segmented in the RFP channel (561 nm) are recognized and marked in red, the nucleus segmented in the GFP channel (488 nm) is recognized and marked in blue, and the overlap between them is recognized and marked with cyan. Scale bar, 5 µm.

Ybr063c (Cnm1) does not affect the extent of ERMES-mediated contacts. (A) ERMES components Mmm1 or Mdm34 were C terminally tagged with GFP on the background of a control strain or strains overexpressing YBR063C (TEF2pr-YBR063C) or deleted for it (Δybr063c). Overexpression of Ybr063c resulted in clustering of ERMES signal to the nuclear ER area but did not change the number or intensity of ERMES puncta. Deleting ybr063c had no effect on these proteins. Scale bar, 5 µm. (B) Ybr063c can be found in distinct areas from ERMES subunits. Overexpressed Ybr063c was N terminally tagged with mCherry on the background of Mdm34 or Mmm1 C terminally tagged with GFP. The yellow arrows represent areas of proximity between the Ybr063c signal and the ERMES proteins, while the white arrows represent areas of Ybr063c signal that does not colocalize with ERMES. Scale bar, 5 µm. (C) A spot dilution assay of strains expressing ybc063c under a GAL promoter in control strains and strains that harbor deletions in mdm34 or vam6. Repressed expression of ybr063c when controlled under the GALpr and grown in glucose caused a complete rescue of the growth defect of Δvam6 in glucose. In contrast, repressing ybr063c on the background of Δmdm34 aggravated the severe growth phenotype of this strain. All strains were grown on both synthetic media with glucose (no expression of Ybr063c) or galactose (Ybr063c is expressed) as a control. (D) 100 representative samples of either the nucleus (on the left) or mitochondria (on the right) that were considered in the quantification analysis of Fig. 3 D. The nuclei were marked by Nsg1-GFP, while the mitochondria were dyed using MitoTracker Orange. (E) Representation of the overlap analysis between the nucleus and mitochondria by artificial intelligence algorithms (ScanR Olympus soft imaging solutions, version 3.2). Mitochondria segmented in the RFP channel (561 nm) are recognized and marked in red, the nucleus segmented in the GFP channel (488 nm) is recognized and marked in blue, and the overlap between them is recognized and marked with cyan. Scale bar, 5 µm.

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