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Fig. S4
Impact of DAPT treatment or gli1/gli3 knockdowns in gene expression pattern and cell proliferation. (A) FISH microscopy of ascl1a and mmp9 in 4dpi retinal sections. (B-C) Low magnification BF microscopy images of mRNA in situ hybridization of mmp9 (B) and ascl1a (C), in DAPT treated retina at 12hpi, showed an increase in its expression as compared with control. (D) Mutated Her/Hes binding sites abolished the impact of nicd over expression on mmp9 promoter, in zebrafish embryo luciferase assay. (E) BF image of mRNA in situ hybridization of zic2b in 4 days post amputated zebrafish fin. (F) Luciferase assay showed that mutations to the Gli-BS abolished the impact of Shh signaling in zic2b promoter. (G) Schematic describing experimental regime of MO injection at the time of injury and electroporation at 4dpi, followed by an i.p. injection of BrdU at 5dpi before euthanasia. (H) IF microscopy images revealed decrease and an increase in BrdU+ cells in gli1 and gli3 knockdowns respectively. (I) Low magnification BF microscopy images of mRNA in situ hybridization of zic2b in DAPT treated retina, at 12hpi. (J) BF images of mRNA in situ hybridization of zic2b in sufu or gli3 knockdown in 4dpi retina. (K) Quantification of ascl1a+, mmp9+ and zic2b+ cells in control and sufu knockdown. (L) RT-PCR analysis of indicated genes’ mRNA levels in uninjured retina, control and gli1 knockdown retina in 2.5dpi. Scale bars, 10 μm (A,B,C,H,I,J) and 500 μm (E). Error bars are SD.