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. 2021 Oct 1;32(20):ar2. doi: 10.1091/mbc.E21-02-0087

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© 2021 Kim, Hlavaty, et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 3: — Targeting Ndel1 to mitochondria results in keratin filament reorganization around these organelles. (A, B) WT keratinocytes were cotransfected with keratin 14-GFP (green) and mito-PAGFP (control) or mito-Ndel1 constructs. Mitochondria were labeled by incubating cells with MitoTracker-Red for 30 min before fixation. mito-Ndel1 induced a reorganization of keratins onto the surface of mitochondria. Scale bar is 10 μm. (C) Z-stack through enlarged mitochondria surrounded by keratin filaments. Red, mitochondria; green, keratin 14-GFP. Scale bar is 5 μm. (D, E) FRAP analysis of keratin 14-GFP in control cell and cells transfected with mito-Ndel1. Scale bars in D and E are 10 μm. (F) Desmoplakin null keratinocytes were cotransfected with keratin 14-GFP (green) and mito-Ndel1 constructs. (G) Lis1^fl/fl keratinocytes were incubated with adenoviral-Cre recombinase for 24 h and then cotransfected with keratin 14-GFP (green) and mito-Ndel1 constructs. (H, I) Ndel1 localization in WT (H) and Lis1 null (I) keratinocytes was analyzed using an immunofluorescent stain against Ndel1.