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. 2019 May 15;15(5):e1008165. doi: 10.1371/journal.pgen.1008165

Fig 9. Hdac1 promotes transcriptional repression at RAREs within the ripply3 promoter.

Fig 9

(A) Schematic of DR1 and DR4 RAREs within the ripply3 promoter. (B) EMSAs with RARs for the ripply3 DR1 and DR4 sites. The positive control is a DR5 site found in promoters of other direct targets [133]. (C) ChIP-qPCR for induced GFP-VP16-RARab at the DR1 and DR4 sites. (D-F) ChIP-qPCR for HDAC1, H3K27ac, H3K27me3 at the ripply3 DR1 and DR4 sites in WT sibling and crg mutants. Fold enrichment for C-F was normalized versus IgG pull-down. (G) Deletions of the ripply3 promoter DR1 and DR4 sites using multiplexed gRNAs and Cas9. Asterisks indicate deletions, which were confirmed with Sanger sequencing. (H) RT-qPCR for ripply3 expression in WT sibling and crg mutant embryos with ripply3 DR1 and DR4 promoter deletions at 36 hpf. The control gRNAs target a region ~100kb away from the ripply3 promoter. (I) Model depicting Hdac1 and RAR function regulating ripply3 in SHF development.