Fig. 3

Generation and characterization of transgenic FS-hRPE cells. A Representative bright field and mCherry images of hESC-derived hRPE cells. Images acquired at 20× magnification. B RT-qPCR against a panel of RPE cell markers. Gene expression relative to YWHAZ and normalized to GAPDH. C Representative flow cytometry analysis plots of TYRP-1 and mCherry expression in hESC-derived hRPE (n = 2–3). D ZO-1 immunocytochemistry analysis of hESC-derived hRPE (n = 3–4, scale is 100 µm). E Western blotting analysis of hRPE conditioned media—two different biological replicates of media from each CX3 clone were loaded. sCX3CL1 was identified by C-terminal 8× -His tag (n = 2–3). Full length western blot represented in Additional file 4: Fig. S3. F Quantification of sCX3CL1 secretion by hRPE by assaying supernatant via ELISA specific to CX3CL1 (n = 3 biological replicates, each consisting of 2 technical replicates). One-way ANOVA and Tukey’s post-hoc analysis performed for data in panels A and E. ND, not detected; ns, not significant. **p < 0.01, ***p < 0.001, ****p < 0.0001