Extended Data Fig. 4: Detailed representation of the cryo-SXT workflow and interactions between mitochondria and cytoplasmic PQC compartments. | Nature Cell Biology

Extended Data Fig. 4: Detailed representation of the cryo-SXT workflow and interactions between mitochondria and cytoplasmic PQC compartments.

From: Nuclear and cytoplasmic spatial protein quality control is coordinated by nuclear–vacuolar junctions and perinuclear ESCRT

Extended Data Fig. 4

(a) Optical path through the specimen. Key: COL, cryogenic objective lens; SS, specimen stage; SP, specimen port; MG, motorized goniometer; CIM, cryogenic immersion fluid; CCL, low magnification cryogenic objective; CS, cryogenic specimen; CIE, cryogenic imaging environment; AP, adapter port; AW, a heated, angled anti-reflection window. (b) Alignment of fluorescence and soft x-ray tomographic data using fiducial markers. (c) A representative confocal image of the spatial relationship between the INQ and nucleolus. NLS-LuciTs (INQ) is shown in green, nucleolus in gold, and Hoechst counterstain in blue. Scale bar is 1μm. (d) The interaction between mitochondria and cytoplasmic inclusions is also seen by fluorescence confocal microscopy in a representative image of a cell co-expressing mito-GFP and NES-RFP-LuciTs. NES-LuciTs is shown in purple, mitochondria in cyan, and Hoechst counterstain in blue. Scale bar is 1μm. (e) Representative confocal fluorescence microscopy images taken of WT, fission mutants (dnm1Δ and fis1Δ) and fusion mutant (fzo1Δ and ugo1Δ) cells expressing mito-GFP and NES-DsRed-LuciTs after 120 minutes at 37 °C and treated with 100μM MG132. Mito-GFP is shown in cyan, NES-LuciTs in purple, and Hoechst counterstain in blue. Scale bars are 1μm.

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