Figure 6
Characterization of the Tal1 Δ/Δ inducible 3TFs embryonic stem cell line. (A) 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 clones C3.17 (allele with the shorter deletion) and C3.6 reconstructed based on the sequencing results. (B) Representative flow cytometry analyses of untreated day 1.75 hemangioblast cultures comparing the profile of unstained cells (top) and cultures stained for VE-CAD and CD41 (bottom). 3–4% of the cells in hemangioblast cultures were auto-fluorescent in the red (VE-CAD) channel. (C) Experimental layout used to characterize the mutant line. (D) Top: representative microscopic images of the indicated conditions. VSM cells appear as large wide-spread cells; Endo cells appear as elongated cells; HPs appear as round floating cells. Unt Unt = untreated; Dox Unt = doxycycline added at EB day 2; Dox Dox = doxycycline added at EB day 2 and at day 0 of hemangioblast culture. Bottom: box plots showing the relative frequencies of VSM, Endo, Pre-HPC and HP populations of HE culture from the indicated conditions (n = 3). Significance was determined by Analysis of Variance (ANOVA) test followed by Tukey HSD test for multiple test correction. Error bars correspond to standard deviations.
