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Files in this Data Supplement:
Fig. S1. No increase in cell proliferation or altered apoptosis is seen in SoxB1-transfected RPE cells. (A-C) Confocal images of a section of a BrdU-labeled electroporated eye stained for BrdU (red) and Sox2 (green), with DAPI-labeled nuclei in blue. (D,E) A TUNEL (red)-stained section of a Sox3/LacZ-transfected eye. Transfected cells were visualized with anti-β-Gal (green). E is a high magnification image of the boxed region in D. Scale bars: 10 µm in A; 100 µm in D. le, lens vesicle; nr, presumptive NR; rpe, presumptive RPE.
Fig. S2. Lens and neural crest markers are not affected in SoxB1-transfected optic cup. (A) δ-crystallin expression in a Sox2/LacZ-transfected optic cup at stage 19, as detected by section in situ hybridization. A strong signal was detected in the lens (le) but not in the optic cup, including the thickened transfected areas in the presumptive RPE (arrowheads). (B) A sister section of A immunostained for β-Gal (green), with DAPI-labeled nuclei in blue. (C-F) A Sox2/LacZ-transfected optic cup at stage 18, showing no evidence for neural crest-like development. Transfected regions (β-Gal; green) have intact basal lamina (C,D; laminin; red) and show no expression of the neural crest cell marker HNK-1 (E,F; red). D and F are high magnification images of the boxed regions in C and E, respectively. (G,H) Sox9/LacZ-transfected optic cup undergoes ectopic neural crest-like development. (G) Ectopic delamination of transfected cells (X-Gal staining; blue). H is a high magnification image of the boxed region in G. (I-L) Delamination of transfected cells is associated with the disruption of laminin (red)-positive basal lamina (arrow in J) and the ectopic expression of HNK-1 (red; K,L). J and L are high magnification images of the boxed regions in I and K, respectively. Scale bars: 100µm. le, lens vesicle; nr, presumptive neural retina; rpe, presumptive retinal pigmented epithelium.
Fig. S3. Expression patterns of transcription factor genes. (A-R) Transverse sections through the eye primordium at stage 10-11 (A,D,G,J,M,P), stage 12-13 (B,E,H,K,Q), stage 14 (N) and stage 16 (C,F,I,L,O,R), stained by in situ hybridization for transcripts of Mitf (A-C), Otx2 (D-F), Six3 (G-I), Rx1 (J-L), Chx10 (M-O) and Optx2 (P-R). Scale bar: 100 µm. le, lens vesicle; nr, presumptive NR; ov, optic vesicle; rpe, presumptive RPE.
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