Rpe65

The eyes of pigs were fixed in 4% paraformaldehyde for 1 week, embedded in paraffin, sectioned, and stained. Sections stained with H&E were used to observe the structure of the eye. The samples were stained with MITF antibody (1:500; Abcam, Cambridge, MA), PAX6 antibody (1:50; Abcam, Cambridge, MA), RPE65 (1:500; Santa Cruz Biotechnology Inc., CA, USA), ZO‐1 (1:500; Invitrogen, California, MA), K12 (1:100; Santa Cruz Biotechnology Inc., California, USA), and Bestrophin antibody (1:200; Abcam, Cambridge, MA) according to the manufacturer's protocol overnight at 4°C. The second antibodies used an Alexa Fluor 594 goat anti‐mouse IgG (1:200) and an Alexa Fluor 488 goat anti‐rabbit IgG (1:200) according to the manufacturer's protocol (ZSGB‐Bio, Beijing, China) for 1 h at RT. After antibody treatments, sections were mounted in H33342 (Vector Laboratory Inc., Burlingame, CA, USA) for nuclear counterstaining and examined by confocal laser scanning microscopy using an LSM 5 PASCAL (ZEISS, Oberkochen, Germany).

BMSCs were fixed with 4 % paraformaldehyde at days 7 or 14 post-neural induction, blocked in 1 % bovine serum albumin and 0.2% Triton X-100 (Sigma-Aldrich, MO, USA), and incubated at 37°C for 1h with primary antibodies against glial fibrillary acidic protein (GFAP; 1:200; rabbit anti mouse; Santa Cruz, CA, USA), nestin (a neural stem cell marker; 1:200; Sigma, goat anti mouse; MO, USA), neuron-specific nuclear protein (NeuN; retinal ganglion cell marker; 1:200; rabbit anti mouse; Abcam, MA, USA), RPE65 (an RPE marker; 1:200; rabbit anti mouse; Santa Cruz, CA, USA) and rhodopsin (a photoreceptor marker; 1:200; rabbit anti mouse; Abcam, MA, USA). The cells were subsequently incubated at 37°C with fluorescein-conjugated secondary antibodies (Dako, CA, USA), mounted and observed under a fluorescence microscope.