Figure S3.

(A, B) The promoter schematics (upper) of ascl1a (A) and oct4 (B) genes reveal the presence of Oct4-BSs, which is confirmed to be functional in a ChIP assay (lower) performed in 16 hpi retinal lysates. (C) BF confocal microscopy images of retinal cross sections show decreased ascl1a and oct4 mRNA with oct4 knockdown at 4 dpi. (D, E) The ascl1a promoter activity assayed in zebrafish embryos co-injected with ascl1a:EGFP-luciferase reporter construct and Renilla luciferase mRNA reveals a dose-dependent decrease with oct4 MO (D); *P < 0.004 (t test), N = 4 and increase with oct4 mRNA (E); *P < 0.005 (t test), N = 4, injected conditions. (F) qPCR analysis of cyclins and delta family genes in oct4 MO-electroporated retina, at 2 dpi; *P < 0.04 (t test), N = 4. (G, H) qPCR analysis of snail gene family (G) and cdh1 (H) mRNAs from GFP⁺ MGPCs compared with the GFP⁻ cells present in rest of the retina from 1016tuba1a:GFP transgenic fish at 4 dpi; *P < 0.004 (t test), N = 4. (I, L, M) qPCR analysis of zeb1a, zeb2a mRNA (I); miR-200a, miR-200b (L); and miR-143, miR-145 (M) miRNA levels at various time points post retinal injury. (J) qPCR analysis of zeb gene family mRNAs from GFP⁺ MGPCs when compared with the GFP⁻ ones present in the same retina from 1016tuba1a:GFP transgenic fish at 4 dpi; *P < 0.005 (t test), N = 4. (K) qPCR analysis of cdh1 and oct4 mRNA levels in zeb2a-transfected retina at 2 dpi and 16 hpi, compared with gfp control mRNA transfection. (N) qPCR analysis of miR-143/miR-45 and miR-200a/miR-200b miRNA levels from GFP⁺ MGPCs compared with the GFP⁻ cells present in rest of the retina from 1016tuba1a:GFP transgenic fish at 4 dpi; *P < 0.0001 (t test), N = 4. Ctl MO is control MO. Error bars are SD. (C) Scale bars, 10 μm; the asterisk marks the injury site, GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer (C).