Supplementary Figures

Files in this Data Supplement:

• Supplementary Figure S1 - (PDF)
  Phase contrast light microscopic images of RPE cultures in SFRM-B27. (A) RPE:choroid explant culture 5 days after initial adherence to laminin coated plastic, demonstrating the difference in appearance between an expanding RPE monolayer (lower portion of the panel) and a multilayered colony of contaminating choroidal fibroblasts (upper portion of the panel). Arrows point to the well-demarcated boundary between the cell types. (B) RPE cells visualized on the surface of a pigmented spheroid grown in suspension culture (prior to initial platedown). (C) Low power image of a T-75 flask containing fifth passage RPE established from pigmented spheroids (method 3) and maintained in culture for nine months. (D) High power image of the RPE monolayer culture shown in panel C. Low (E) and high (F) magnification images of RPE cells continuing to emanate from a re-used pigmented spheroid after being plated onto laminin coated plastic a third time. Scale bars: 50 mm (panels A, B and D) and 100 mm (panel E).

• Supplementary Figure S2 - (PDF)
  The relative fold change in the expression of bestrophin was determined across five individual RPE cultures by quantitative PCR. Culture 1 was arbitrarily designated as the reference culture, and results are expressed as the mean ± S.E.M. of three separate amplifications. The method of origin, age in culture, passage number and spheroid platedown number (applicable for method 3 only) for each culture is provided in the table below the graph. N/A = not applicable.