Figure 2 | Scientific Reports

Figure 2

From: Endogenous suppression of WNT signalling in human embryonic stem cells leads to low differentiation propensity towards definitive endoderm

Figure 2

Micropatterned differentiation confirms the low differentiation propensity of VUB04 towards definitive endoderm (DE). (A) Schematic overview of micropatterned differentiation protocols used to direct four hESC lines towards primitive streak and DE like fates in 1000 μm in diameter circular colonies. Created with BioRender. (B) Representative immunofluorescent images for T and SOX2 after 48-h micropatterned differentiation towards primitive streak fates. (C) Percentage of T- and SOX2-positive cells after peri-gastrulation-like differentiation observed within the tested lines. Each data point represents one individual colony. Number of colonies were 120, 91, 120 and 134 for VUB02, VUB01, VUB03 and VUB04, respectively. Data pooled from two independent experiments. Error bars represent mean ± SD. (D) Representative images for SOX17 and FOXA2 after 48-h DE micropatterned differentiation. (E) Percentage of SOX17-positive, FOXA2-positive and double-positive cells after DE differentiation observed in the tested lines. Each data point represents one individual colony. Number of colonies were 370, 318, 201 and 300 for VUB02, VUB01, VUB03 and VUB04, respectively. Data pooled from two independent experiments. Error bars represent mean ± SD. All p-values were calculated using one-way ANOVA. All scale bars represent 100 μm.

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