Supplementary Figure 7: Supporting results for the genetic targeting of KLF4 binding site within the Tbx3 hub (Related to Figure 7).

a, IGV tracks of H3K27ac and KLF4 ChIP-seq in PSCs showing the whole Tbx3 distal enhancer (top), the region that was deleted by CRISPR/Cas9 (Dist-KO, bottom, see Fig. 4f) and the ___location of the gRNA used to mutate a specific KLF4 binding motif (Dis-KLF4mut gRNA). b, Genotyping strategy of the surveyor assay used to detect mutation/indel at the target KLF4 binding site within the distal Tbx3 enhancer (Dis-KLF4mut). The results for 4 homozygous mutant clones (mut1-4) are shown (See also Supplementary Fig. 8). c, Sequencing results of the four mutant (mut) clones compared to the wild type (WT). d, ChIP-qPCR showing the relative levels of KLF4 binding to Tbx3 distal enhancer in n = 2 WT clones and n = 4 mut clones (left panel). Values show ChIP signal over input. As control, binding of KLF4 to an unaffected region (Fbxo15 promoter) was tested (right panel). Unpaired on-tailed t-test was performed to compare WT vs MUT and the specific p-values are shown in the graph.