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. 2024 Apr 30;121(19):e2317703121. doi: 10.1073/pnas.2317703121

Fig. 4.

Fig. 4.

PKMO FX is compatible with various labeling strategies to achieve multicolor imaging of fixed cells. (A) Multicolor labeling strategies for imaging of mitochondrial subcompartments and related organelles. Abbreviations: IM (inner mitochondrial membrane); OM (outer mitochondrial membrane). (B) Two-color STED images of HeLa cells transfected with HaloTag-TOM20 and labeled with CA-SiR for OM and PKMO FX for IM. From Left to Right: split channels of IM, OM, and composite. (Scale bars, 2 μm.) (C) Two-color STED and confocal images of HeLa cells transfected with Lifeact-EGFP and stained with PKMO FX. Left: zoomed-in images of the white boxed areas of IM and actin. Actin was recorded in the confocal mode. Right: composite. (Scale bars, 2 μm.) (D) Two-color STED images of HeLa cells costained with PKMO FX and a fixable DNA probe: SiR-DNA. Left: zoomed-in images of the white boxed areas of IM and DNA. Right: composite. (Scale bars, 2 μm.) (E) Two-color confocal images of HeLa cells costained with PKMO FX and a fixable actin probe: SPY650-FastAct. (Scale bar, 5 μm.) (F) Flowchart showing the experimental procedure of the EdU-based metabolic labeling. (G) Two different cell states demonstrated by metabolic labeling. Left: cell cycle state I in which both nascent nuclear DNA (nDNA) and nascent mitochondrial DNA (mtDNA) are present. Right: cell cycle state II in which only nascent mtDNA spots are present. (Scale bars, 5 μm.)