Figure 4

5TFs overexpression in hemangioblast cultures cannot generate VE-CAD⁺CD41⁺ cells in the absence of a functional TAL1. (A) CRISPR/Cas9 strategy used to disrupt the Tal1 gene in the i5TFs mESC line (figure downloaded and modified from the Ensembl genome browser). (B) Left: Electrophoresis showing the genotyping of the Tal1 mutant mESCs clones used for experiments. Right: Scheme of the WT Tal1 gene and the Tal1 gene from mutant clone C3.39 reconstructed based on the sequencing results. (C) Representative microscopy images of the indicated conditions at day 3 of hemangioblast culture. (D) Representative flow cytometry analyses of the indicated conditions (left panels) and box plots showing the relative frequencies of VSM, V⁺CD⁻, V⁺CD⁺ and HP populations at day 3 of hemangioblast culture (n = 3). Significance was determined by Analysis of Variance (ANOVA) test. Error bars correspond to standard deviations.