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. 2011 Mar 21;52(3):1573–1585. doi: 10.1167/iovs.10-6413

Figure 2.

Figure 2.

RT-PCR, Western blot analysis, and immunofluorescent staining of RPE-specific genes expressed by hES-RPE derived from hES-3 embryonic stem cell line and control cells. Polarized hES-RPE (p-hES-RPE; passage 3) and polarized fetal human RPE (p-fRPE) were compared with nonpolarized hES-RPE (Np-hES-RPE; passage 3), nonpolarized fetal human RPE (Np-fRPE), nonpolarized ARPE-19 (Np-ARPE), and undifferentiated hES3 cells. The polarized hES-RPE expressed high levels of RPE-specific gene products. (A) RT-PCR for CRALBP, PEDF, RPE65, and VMD2; GAPDH was the loading control. (B) Western blot for PEDF and RPE65 with GAPDH as loading control. (C–F) Immunofluorescent staining. PEDF was highly expressed in polarized hES-RPE cells at passage 3 (C) and less prominently expressed in nonpolarized hES-RPE cells (E). Immunofluorescent staining for RPE65 was found in polarized RPE cells (D), but not in nonpolarized hES-RPE cells (F). Immunoreactivity was detected using secondary antibodies conjugated to FITC (green). Nuclei are counterstained with DAPI (blue).