Figure 2.

Figure 2.

(A) An experimental timeline that describes the MO delivery, electroporation, and BrdU pulse before harvesting at 4 dpi. (B, C) IF confocal microscopy images of retinal cross sections show the decline in BrdU⁺ MGPCs with increasing concentrations of oct4 MO (lissamine tag) at 4 dpi (B), which is quantified (C); *P < 0.0001 (t test), N = 4. (D) An experimental timeline that describes the MO delivery, electroporation, and harvest at 16 hpi and 2 dpi. (E) The qRT-PCR analysis of oct4, ascl1a, and sox2 genes in oct4 knockdown retina at 2 dpi and 16 hpi; *P < 0.01 (t test), N = 4. (F) Western blot analysis of Oct4, Ascl1a, and Sox2 from retinal extracts collected after oct4 knockdown at 16 hpi and 2 dpi. Gapdh is the loading control. (G, H) RT-PCR (G) and qRT-PCR (H) of lin28a and her4.1 in oct4 knockdown retina at 2 dpi. (I) BF microscopy images of retinal cross sections show the expression of lin28a and her4.1 mRNA in oct4 knockdown retina at 4 dpi. (J) The her4.1 promoter schematic reveals the typical Oct4-BS (upper) and the retinal ChIP assay confirms the physical binding of Oct4 at the typical BS (lower) in 16 hpi retina. Ctl MO is control MO. Error bars are SD. (B, I) Scale bars, 10 μm; the asterisk marks the injury site; GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer (B, I).