Extended Data Fig. 7: Premarking in ES cells is required for robust future macrophage enhancer activation.

a, b, Putative Prdx5 enhancer mutant sequence diagram and DNA sequence documentation of several of the homozygous mutation clones, used for analysis. c, Three mutant clones (B11, F1, G11) of the putative Prdx5 enhancer decreased level of Prdx5 eRNA transcription in ESDMs (right). Each dot indicates one biological experiment (n = 3 biological repeats from two pooled different experiments; n = 2 biological repeats from two pooled different experiments for B11). ESRRB binding in ES cells is a representative experiment (left) (n = 2 biological repeats). d, Nod2 eRNA transcription was tested in wild-type and mutant (B11, F1, G11) clonal cells of the Prdx5 enhancer. Data from one representative experiment are plotted (n = 2 biological repeats). e, f, Putative Nod2 enhancer mutant sequence diagram and DNA sequence documentation of several of the homozygous mutation clones, used for analysis. g, Two mutant clones (D2, D12) of the putative Nod2 enhancer decreased level of Nod2 eRNA transcription in ESDMs (right). Each dot indicates one biological experiment (n = 3 biological repeats from two pooled different experiments; n = 2 biological repeats from two pooled different experiments for D12). ESRRB binding in ES cells is a representative experiment (left) (n = 2 biological repeats). In the box plots, line shows median and box shows 25th and 75th percentiles. P values calculated using Welch’s two-sided t-test. Primer sets are listed in Supplementary Table 2.