Fig. 2.

Enrichment of cardiac VISTA enhancers in cPIRs. a Percentages of cPIRs or hESC PIRs overlapping with VISTA enhancers of the heart, branchial arch and limb. Results obtained with randomised PIRs (1000 permutations) are shown as controls. The p values were generated from permutation tests performed to compare observed vs. randomised PIRs and corrected for multiple testing using Bonferroni procedure. b Image of mm172-reporter staining of an E11.5 mouse embryo obtained from the VISTA enhancer browser (https://enhancer.lbl.gov/)¹⁴. c Promoter–cPIR interactions of hESC-CM occurring between the promoters of TTC29 and EDNRA (thick arc line represents the interaction between EDNRA’s promoter and mm172). d PCR using gDNA/cDNA to detect mm172 deletion/Ednra expression and immunoblotting to detect EDNRA (glycosylated) were performed to compare rat cardiomyoblast H9c2 cell lines transfected with the empty plentiCRISPRv2 plasmid or the same plasmid cloned with two gRNA sequences to target mm172. Actb/ACTB is the loading control