Fig. 6: Homozygous deletion of putative regulatory elements of Notch1 and Pecam1 genes reduced their expression during EC differentiation.
From: Endothelial gene regulatory elements associated with cardiopharyngeal lineage differentiation
a Scheme of the steps of targeted Pecam1- enh.int2 and Notch1- enh.int15 deletion with CRISPR/Cas9. Red lines indicate the position of the two gRNAs used. b Quantitative real-time PCR (qPCR) analysis of Notch1 mRNA expression level in mESC Notch1-∆enh.in15 (clones #7 G; #11B) and Pecam1 in mESC Pecam1∆enh.in2 (clones #7G; 5G) during EC differentiation. Notch1 and Pecam1 expression was reduced in mutant cell lines (in red), compared to WT cells (in black), used as control. The X-axis denotes the three time points (d4-d6-d8); the y-axis indicates the expression level, evaluated using the \({2}^{-\varDelta {Ct}}\) method. Gapdh expression is used as a normalizer. Values are the average of five biological replicates \(\pm\) standard deviation (SD). p value (*) <0.05; p value (**) <0.01 and p value (***) <0.001 are considered significant; ns no statistical significance (parametric paired t-test, one-tailed). c Quantitative real-time PCR (qPCR) analysis of gene expression of Notch1-related genes in mESC Notch1-∆enh.in15 (clones #7G; #11B) during EC differentiation. p value (*) <0.05 and p value (**) <0.01 are considered significant; ns no statistical significance (parametric paired t-test, one-tailed).
