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Increased co-localization of <italic>oct4</italic> with MGPCs towards later stages of retina regeneration, and the effect of <italic>ascl1a</italic> knockdown on <italic>oct4</italic> expression.(A, B) BF and IF confocal microscopy images of retinal cross sections show increased oct4 mRNA in BrdU+ MGPCs towards the later stages of retina regeneration (A), which is quantified (B); *P < 0.0001 (t test), N = 4. (A) White arrowheads mark BrdU+ cells and white arrows mark oct4+ cells in (A). (C) IF confocal microscopy images of retinal cross sections from 1016tuba1a:GFP transgenic retina show the GFP expression in PCNA+ MGPCs at 4 dpi. White arrows mark co-labeled PCNA and GFP expression. (D) RT-PCR (upper) and qPCR (lower) analysis of oct4 mRNA in ascl1a knockdown retina at 2.5 dpi. (E) BF microscopy images of retinal cross sections show the decline in oct4 mRNA with increasing concentrations of ascl1a MO at 4 dpi. (F) The oct4 promoter schematic reveals the typical Ascl1a-BSs (upper) and the retinal ChIP assays confirm the physical binding of Ascl1a at the typical BSs (lower) in 16 hpi retina. Ctl MO is control MO. Error bars are SD. (A, C, E) Scale bars, 10 μm; the asterisk marks the injury site, GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer (A, C, E).
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