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. 2019 Feb 4;218(2):489–507. doi: 10.1083/jcb.201802113

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© 2019 Mitra et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 7. — Mycb regulates her4.1 gene transcription in injured retina. (A and B) RT-PCR (top) and qPCR (bottom) show increased her4.1 induction with MO-based mycb knockdown (A) or 10058-F4 (B) relative to control MO and DMSO, respectively, in 2-dpi retina. *, P < 0.005. UC, uninjured control. (C) FISH and IF microscopy shows that 10058-F4 treatment increases her4.1 expression compared with water or DMSO-treated control in 4-dpi retina. Bar, 10 µm. White asterisks mark the injury sites. (D) Myc inhibition through 10058-F4 up-regulates her4.1:gfp-luciferase expression compared with control and DAPT-treated embryos. (E) Diagram of her4.1 promoter with putative Mycb-binding sites. The solid lines represent DNA sequences of the promoter. (F) The retina ChIP assay at 4 dpi reveals Myc and Hdac1 bound to Myc-BS on her4.1 promoter. (G) qPCR analysis of her4.1 mRNA from GFP⁺ and GFP⁻ MGPCs sorted from 1016 tuba1a:gfp transgenic fish retina with 1 µM 10058-F4 treatment at 4 dpi, compared with WT. n = 3 biological replicates in all experiments. ONL, outer nuclear layer; INL, inner nuclear layer; N.S., nonspecific; BS, binding site.