Figure 11. TdTomato expression and RPE65 antibody staining in cryosections. The images show coexpression of endogenous Rpe65 protein (green) and tdTomato (red) in RPE cells following subretinal injection and electroporation of the pVAX-tdTomato plasmid. A: Shown is the red channel. B: Illustrated is the green channel. C: Shown is a composite of the green red and blue (DAPI stained nuclei) channels. D: The red and green colocalized in the RPE cell layer (yellow) but not completely. E: There were some areas where the green of the Rpe65 protein was observed more apically in the RPE cells and the red of the tdTomato protein was more basolateral and perinuclear. A white arrow drawn across the panel represents an in silico section through which the red and green channels were quantified. F: Shown are the relative intensities of red (tdTomato), green (Rpe65), and blue (DAPI) fluorescence in a plot. The intensities follow the white arrow drawn through several RPE cells to scale below the image in E. This confocal image was obtained from a mouse eye harvested four days after injection. The RPE cells have survived treatment and coexpress Rpe65 protein and tdTomato in the same cell, though in slightly different locations within the cell. The Rpe65 protein accumulated more on the apical top half of the cell, while the tdTomato accumulated more basally and more in the perinuclear region. The scale bar represents 50 μm.