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. 2019 Jul 15;132(14):jcs232009. doi: 10.1242/jcs.232009

Fig. 4.

Fig. 4.

Functional mitochondria are required for mitochondrial Rhod-2 transfer to melanosomes. (A–F) Confocal immunofluorescence images of melan-a melanocytes transfected with mito-GFP and labeled with Rhod-2 in the presence of DMSO (control) or 10 μM CCCP, as indicated. All the time points are time after washing with the indicated medium following dye loading for 30 min. Insets show 6× magnified images of the boxed region. Scale bar: 10 μm. (G–I) Confocal immunofluorescence images of melan-a melanocytes transfected with MART1–GFP and labeled with Rhod-2 in the presence of DMSO (control) or 10 μM CCCP, respectively, 60 min after dye loading (for 30 min) and washing with indicated medium. Insets show 6× magnified images of the boxed region. Scale bar: 10 μm. (J–L) Confocal immunofluorescence images of melanocytes transfected with mito-GFP then labeled with Rhod-2 in the presence of indicated compounds (60 min after washing with indicated medium following dye loading for 30 min): DMSO (J); oligomycin 10 μM (K); oligomycin 10 μM washout (L). Arrowheads, Rhod-2 dots separated from mito-GFP. Insets show 6× magnified images of the boxed region. Scale bar: 10 μm. (M) Quantification of the number of Rhod-2 dots in a single cell (60 min after wash) for each treatment; n=245, 170, 149, 123, 117, 130, and 107 cells, respectively. **P<0.01; ***P<0.001 (one-way ANOVA).