Fig. 3. Expression of the µ1A subunit in melanocytic cells. (A) µ1A transcript levels determined by real-time PCR. Note the expression of µ1A in 1106c normal human melanocytes was higher than in other melanocytic cells or in NHEK. Values are mean ± s.d. of two experiments. (B) Melanoma cells as noted were transiently transfected with a plasmid containing µ1A tagged with HA, or without a plasmid as a control (Ctrl). Comparison of AP1 and µ1A-HA subunits was performed by immunoblotting in those cell lines. Positions of molecular size markers are indicated on the left in kDa. (C-H) Melanoma cells as noted were fixed 48 hours after transfection with µ1A-HA, and stained with a combination of PEP13h (red) and anti-HA (green) antibodies. Specimens were analyzed by confocal microscopy, and yellow fluorescence indicates colocalization. Nuclei were counterstained with DAPI (blue). Representative images are from at least two independent experiments. Bars, 10 µm (C,F); 1 µm (D,E,G,H).