Extended Data Fig. 8: Validation of studied ScMrs2 forms in terms of cellular localization and expression.
WT represents the exploited S. cerevisiae strain (BY4741) used for the experiments. ΔScMrs2 is BY4741 lacking native Mrs2. P426-ADH-HA(N) is ΔScMrs2 transformed with empty vector. ScMrs2_Full length is ΔScMrs2 transformed with the P426-ADH-HA(N) vector harbouring full−length GFP-tagged ScMrs2, forming the basis for structure-function analyses. The corresponding variants of CtMrs2 are indicated in bold. a, The localization of ScMrs2 wild-type (ScMrs2_Full length) and mutants were investigated using fluorescence microscopy, comparing GFP-fluorescence (left), differential interference contrast (DIC) (middle) and merged (right), the scale bar represents 5 µm, Imaging experiments were conducted for three times with different clones of each yeast strain, and each time five different regions were captured containing around 100 cells in total. b, The expression of ScMrs2 wild-type (ScMrs2_Full length) and mutants probed using immunoblotting with an anti-HA antibody and using phosphoglycerate kinase (Pgk1) as a loading control. The experiments were conducted three times with different clones of each yeast strain.
