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. 2022 Jun 29;221(8):e202103048. doi: 10.1083/jcb.202103048

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© 2022 Bisinski et al.

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Figure S3. — Cvm1 does not co-localize with markers of the lipid droplets, endosomes, the Golgi complex, the trans-Golgi network, or the plasma membrane. Related to Fig. 4. (A–D) Fluorescence microscopy analysis of the localization of GFP-Cvm1 under the control of the NOP1 promoter with markers of different organelles. Erg6-mKate2 was used as a marker for lipid droplets, Vps8-Halo as a marker of late endosomes, Sec7-Halo as a marker of the trans-Golgi network/early endosomes, and Mnn9-Halo as a marker of the early Golgi complex. All strains containing a Halo-tagged protein were labeled with the JF646 ligand. Lipid droplets were imaged in logarithmic and stationary phase, because their morphology differs in these two growth phases. No significant colocalization was observed between GFP-Cvm1 and any of the markers. Scale bars represent 2 µm. BF = Brightfield. (E) Fluorescence microscopy analysis of the localization of mCherry-Cvm1 under the control of the TEF1 promoter and Pma1-GFP as a marker of the plasma membrane. No significant colocalization was observed between the two signals. Scale bar represents 2 µm. (F) Colocalization of Cvm1 with Vac8. Fluorescence microscopy images of a strain expressing GFP-Cvm1 under the control of the NOP1 promoter and Vac8-mKate2. Both proteins localize along the vacuole membrane. Some regions of enrichment of Cvm1 are also enriched in Vac8 compared with the rest of the vacuole membrane. Scale bar represents 2 µm.