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

Figure 9.

Figure 9.

Apoptosis (TUNEL) assay to measure RPC death in various CM after overnight challenge with 250 μM H2O2. The RPCs cultured in the polarized hES-RPE (passage 3) CM, polarized fRPE CM, or RPC medium with 50 ng/mL PEDF supplement showed less cell death than those cultured in the regular RPC medium, HEK-CM, polarized fRPE CM, or hES-RPE CM with anti-PEDF antibody. Images (A–H) Fluorescent microscopic images of TUNEL+ RPCs. (A) RPCs cultured in the regular RPC medium; (B) RPCs in HEK-CM; (C) RPCs in RPC medium with 50 ng/mL PEDF; (D) RPCs in RPC medium with 50 ng/mL PEDF and 20 μg/mL anti-PEDF antibody; (E) RPCs in the polarized fRPE CM; (F) RPCs in the polarized fRPE CM with 20 μg /mL anti-PEDF antibody; (G) RPCs in the polarized hES-RPE CM; (H) RPCs in the polarized hES-RPE CM with 20 μg/mL anti-PEDF antibody. (I) The quantification of apoptotic (TUNEL+) RPCs from three individual experiments. The RPCs cultured in the polarized hES-RPE CM, polarized fRPE CM, or RPC medium with recombinant PEDF supplement had significantly fewer apoptotic cells than those cultured in the regular RPC medium, HEK-CM, and in the polarized fRPE CM, hES-RPE CM, and recombinant PEDF with anti-PEDF antibody; **P < 0.01.