Figure S4.
Subcellular localization of ERMES fusion proteins. Representative images of localization of mNG-fused proteins. Kar2(1–45)-mScarletI-KDEL (SP-mScI-KDEL) and Su9(1–69)-mTurquoise2 (Su9-mTq2) are ER and mitochondrial markers, respectively. Scale bars are 2 μm. Refer to the image caption for details. The seven-row fluorescence microscopy montage evaluates the subcellular distribution of various mNeonGreen (mNG)-tagged constructs in a quadruple ERMES deletion strain. The layout consists of four columns: the mNG signal, ER marker (SP-mScl-KDEL), mitochondrial marker (Su9-mTq2), and a multi-channel Merge. The top three rows display standard fusions (MMM1-mNG, MDM12-mNG, MDM34-mNG), while the subsequent rows feature synthetic tethering constructs designed to bridge organelles. Specifically, MMM1-mNG-FIS1C, SEC66N-MDM12-mNG-FIS1C, and SEC66N-MDM34-mNG-FIS1C show concentrated signals at membrane interfaces, whereas the MMM1(mut)-mNG-FIS1C variant exhibits altered localization. Dashed lines outline individual cell boundaries in the merged view, and scale bars are 2 micrometers.

Subcellular localization of ERMES fusion proteins. Representative images of localization of mNG-fused proteins. Kar2(1–45)-mScarletI-KDEL (SP-mScI-KDEL) and Su9(1–69)-mTurquoise2 (Su9-mTq2) are ER and mitochondrial markers, respectively. Scale bars are 2 μm.

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