Fig. 1

Similarities and differences between non-pigmented rabbit RPE cells and CECs in situ. a Schematic diagram of the morphology and location of RPE cells and CECs in rabbits created with BioRender.com. b qRT-PCR analysis of OTX2, CRELBP, BEST1, MITF, RPE65, ATP1A1, and TJP1 between Rb-RPE and Rb-CEC. Quantification represented the levels of relative mRNA expressions normalized to GAPDH. c Representative immunofluorescent images of Rb-RPE and Rb-CEC in situ were collected, including ZO1 (green), ATP1A1 (red) and retinal pigment epithelial markers MITF (green), and RPE65 (red). Nuclei were stained with DAPI (blue) (scale bar: 50 μm). d Cell density, hexagonality, and coefficient of variation analysis were based on ZO1 immunostaining. e Scanning electron microscope showed a regular hexagonal shape in both Rb-RPE and Rb-CEC, which were well formed and with distinct cell boundaries (scale bar: 10 μm) (Upper). Transmission electron microscope showed that both types of cells were attached to their respective basement membrane, named BM and DM, by hemidesmosomal junctions (▲) (scale bar: 500 nm) (Middle). Adjacent cells were joined with numerous well-developed tight junctions (▲) (scale bar: 500 nm) (Lower). Data are mean ± SEM. All results were obtained from three independent experiments. Significance (*P < 0.05, **P < 0.01 and ns: nonsignificant) relative to Rb-CEC. RPE, retinal pigment epithelium; CEC, corneal endothelial cell; qRT-PCR, quantitative real-time reverse transcription polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; BM, Bruch’s membrane; DM, Descemet’s membrane; Rb-CEC, rabbit CECs; Rb-RPE, rabbit RPE cells; SEM, standard error of the mean