Extended Data Fig. 2: Overexpressed Mge1 localizes to both mitochondria and the nucleus.
From: A direct role for a mitochondrial targeting sequence in signalling stress
(a) Live-cell fluorescence imaging of cells expressing the mitochondrial marker Tom70-GFP and either Mge1-mCherry (top) or Mss116-mCherry (bottom). Overexpression of MGE1-mCherry and MSS116-mCherry from the GAL1-10 promoter was induced by the addition of galactose for 4 h. Zoom-in images of representative cells are shown on the right. Scale bars, 5 μm. Mean Pearson’s correlation coefficients is shown on the right (n = 3 independent experiments; in total of 60 cells per strain; Two-tailed Student’s t-test). (b) Large fields of cells analyzed in Fig. 1e, overexpressing Mge1-mCherry (top) or Mss116-mCherry (bottom) as well as the nuclear envelope marker Nup159-GFP. Representative zoom-in images and corresponding fluorescence intensity profiles along the white line are shown on the right. Scale bars, 5 μm. (c) Large fields of cells analyzed in Fig. 1f, expressing the nuclear protein GFP1–10-mCherry-Pus1 alone or with either Mge1-GFP11 or Mss116-GFP11. Zoom-in images of representative cells are shown on the right. Scale bars, 5 μm. Overexpression of MGE1-GFP11 and MSS116-GFP11 was confirmed by measuring their mRNA levels relative to control cells (bottom panels). n = 3 biological replicates; two-tailed Student’s t-test. (d) CIS1 mRNA levels in cells overexpressing MGE1, ILV2, HEM15 or MRP51 from the GAL1-10 promoter for 4 h (left panel; one-way ANOVA followed by Dunnett’s test). Overexpression was confirmed by qPCR analysis (right panels; two-tailed Student’s t-test). n = 3 biological replicates. EV, empty vector; OE, overexpression. (a, c, d) Data represent mean +/− SD; ns, not significant.
