Figure 2.

Microphotographs of the immunolabeling of newborn pig ciliary epithelium (CE)-derived cells after in vitro differentiation on poly-D-Lysine, laminin coated coverslips in the presence of 1% serum and 10 ng/ml basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). The images were acquired using an epifluorescent microscope. A, B: Cells labeled for recoverin are clustered together (arrows). C: 4',6-diamidino-2-phenylindole (DAPI) nuclear staining corresponding to A and B. D, E: Cells double-labeled for recoverin (red, D) and rhodopsin (green, D and E) are depicted by arrows. The focus is set to show rhodopsin-positive cell processes. Strong recoverin staining in the cytoplasm masks the nuclear DAPI staining, which is shown separately in F. Cells positive for protein kinase α (PKCα; green in G and H, arrows) did not co-label with recoverin (red in G and H and another example in the inset in G, arrowheads). The focus is set to show the processes of PKCα-labeled cells in G and H, and the recoverin-labeled processes in the inset in G. Corresponding DAPI nuclear stain is shown in I. J-K: A calbindin immunopositive cell is depicted by the arrow. The focus is set to show the processes of the labeled cell. Corresponding DAPI nuclear stain is shown in L. M, N: Recoverin-positive cells (arrows in M and N) that had retained pigmented granules (arrow in the bright-field image in O). P, Q: RPE-65 immunopositive cells (arrows) and the corresponding nuclear DAPI staining in R.