Panel A: The panel shows a series of images depicting the subcellular localization of melanosomes in RAB27A-deficient melanocytes expressing different RAB27A variants. The images include EGFP fluorescence, bright-field images, and merged images. The EGFP-tagged RAB27A variants are shown in green, and melanosomes are pseudo-colored in red. Panel B: The panel shows a bar graph representing the percentage of cells with peripheral melanosome distribution. The x-axis labels the different conditions (Control, WT, S115R, S106FfsTer18), and the y-axis shows the percentage. Error bars indicate the standard error of three independent experiments. Panel C: The panel shows an immunoblot analysis of RAB27A protein expression in melan-ash cells. The blot is probed with anti-GFP and anti-beta-actin antibodies, with molecular mass markers indicated in kilodaltons.
Subcellular localization of melanosomes in RAΒ27A-deficient melanocytes transiently expressing RAΒ27A variants. (A) EGFP alone (control) or EGFP-tagged RAB27A (WT, Ser115Arg [indicated as S115R], and Ser106PhefsTer18 [indicated as S106FfsTer18]) were transiently expressed in melan-ash cells. EGFP is shown in green, and melanosomes in the right panels (Merge) are pseudo-colored in red. The insets show magnified views of the boxed areas. The arrowheads indicate the colocalization between EGFP-RAΒ27A (WT) and melanosomes. Scale bars, 20 μm. (B) Percentage of cells showing peripheral melanosome distribution is shown in A. Error bars indicate means ± SE of three independent experiments (n > 25 cells for each experiment), which were performed on different days. ***P < 0.001 (one-way analysis of variance and Tukey’s test). (C) Expression of RAB27A proteins in melan-ash cells is shown in A as determined by immunoblotting with the antibodies indicated. The positions of the molecular mass markers (in kilodaltons) are shown on the left. Source data are available for this figure: SourceData F4.