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Fig. 1 | Stem Cell Research & Therapy

Fig. 1

From: MSX2 suppression through inhibition of TGFβ signaling enhances hematopoietic differentiation of human embryonic stem cells

Fig. 1

MSX2 is suppressed upon inhibition of TGFβ signaling during hematopoietic differentiation of hESCs. a Schematic overview of hESC hematopoietic differentiation using a chemically defined system. SB431542 (SB) was added during days 5–8, and RNA-seq was performed on CD31+ cells at day 8. b GSEA of TGFβ signaling-associated gene sets with or without SB treatment. c Heatmap of hematopoiesis-related signature genes with or without SB addition. d GSEA of hematopoiesis-associated gene sets with or without SB treatment. e Schematic diagram showing the strategy of screening potential mediators of TGFβ signaling in hematopoietic differentiation. The red circle represents downregulated TFs upon SB treatment (also see Fig. S1A). The black circle represents potential TFs upstream of SB-repressed genes analyzed by using Enrichr (also see Fig. S1B). f The real-time PCR analysis of MSX2 expression in cells at day 8 of hematopoietic differentiation with or without SB treatment. g The real-time PCR analysis of MSX2 expression in cells at day 8 of hematopoietic differentiation with or without TGFβ1 treatment. Relative expression is normalized to the level (= 1) of Actin. Results are shown as means ± SD (n = 3). h ChIP-qPCR analysis of SMAD2/3-responsive elements on promoters of MSX2 in H1-derived cells. Non-specific IgG was used as isotype control. All values are normalized to that of their corresponding input samples. Results are shown as means ± SD (n = 3). i Real-time PCR analysis of MSX2 expression in undifferentiated hESCs, MEs (APLNR+), HEPs (CD31+CD34+), and HPCs (CD43+) generated from hematopoietic differentiation of hESCs. Relative expression is normalized to the level (= 1) of undifferentiated hESCs. Results are shown as means ± SD (n = 3). **P < 0.01 and ***P < 0.001

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