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. 2021 Feb 19;12:1157. doi: 10.1038/s41467-021-21426-7

Fig. 5. Identification of a regulatory element for TCF15.

Fig. 5

a ATAC-seq profile at the TCF15 locus. Grey boxes indicate putative enhancers identified (TCF15 Enh-1 and TCF15 Enh-2). RARA footprint identified within TCF15 Enh-2. Mutant reporter sequence for TCF15 Enh-2 RARA mutant. b TCF15 Enh-1 (n = 15/15) and c TCF15 Enh-2 (n = 9/9) reporter expression in presomitic mesoderm (Psm), notochord (Nc), somites (So) and lateral plate mesoderm (Lpm). d Combined TCF15 Enh-1/Enh-2 reporter expression (n = 9/9). e Transverse sections of embryo in c immunostained for Citrine showing TCF15 Enh-2 expression in somites and lateral plate mesoderm, white dashed line in c indicates location of section, representative of (n = 4/4). Nuclei stained with DAPI (blue). f TCF15 Enh-2 Citrine reporter with RARA binding site mutation displays lack of expression in somites (n = 6/6). Epigenome engineering using dCas9-Krab with (g) control scrambled sgRNAs resulted in no change (n = 8/9) and (h) sgRNAs targeting endogenous RARA binding site led to loss of TCF15 expression (n = 6/8) as shown by wholemount in situ hybridisation. i Percentage of embryos with normal (blue) or reduced (orange) TCF15 in situ expression after electroporation of control scrambled sgRNA with dCas9-Krab or sgRNAs targeting TCF15 Enh-2 RARA binding site with dCas9-Krab. All scale bars = 500 μm except for e scale bar = 100 μm.