Figure 5.

Figure 5.

(A) An experimental timeline that describes the mRNA transfection and BrdU pulse (for 4 dpi collection) before harvesting either at 2 or 4 dpi. (B, C) IF confocal microscopy images of retinal cross sections show reduced BrdU⁺ MGPCs at 4 dpi in oct4 mRNA transfected condition, compared with gfp mRNA-transfected control retina (B), which is quantified (C); *P < 0.0001 (t test), N = 4. (D) IF confocal microscopy images of retinal cross sections of oct4 mRNA-transfected retina at 4 dpi shows the cells with strong expression of Oct4 having a significant seclusion from PCNA⁺/BrdU⁺ MGPCs. White arrowheads mark BrdU⁺/PCNA⁺ cells and white arrows mark Oct4⁺ cells. (E) Quantification of BrdU⁺ and PCNA⁺ cells from oct4-overexpressed retina. (F) Western blot analysis of Co-IP of Oct4 and Hdac1 in retinal extracts at various time points postinjury probed with anti-Hdac1 antibody. (G) Western blot analysis of Co-IP of Oct4 and Hdac1 in retinal extracts obtained after hdac1 overexpression at 15 hpi and probed with anti-Hdac1 antibody. (H, I, J, K, L) The qRT-PCR analysis reveals the levels of tgfbr1b, tgfb2, tgfbi, smad7 (H), snails (I), cdh1 (J), lin28a (K), and let-7a miRNA (L) in oct4 mRNA-transfected retina at 2 dpi; *P < 0.001 (t test), N = 4. Error bars are SD. (B, D) Scale bars, 10 μm; the asterisk marks the injury site; GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer (B, D).