Fig. 4
R-spondin2 promotes hematopoietic differentiation by augmenting APLNR+ mesodermal cells. a, b Flow cytometry analysis of the percentage of APLNR+ mesoderm cells (a) at day 3 and CD31+CD34+ HEPs (b) at day 5 of differentiation in mAGM-S3 co-culture from H1 cells with or without the treatment of R-spondin2 (20 ng/mL). Results are shown as means ± SD (n = 3). **P < 0.01. c Heatmap showing expression change of genes associated with pluripotency and three germ layers after R-spondin2 treatment (log10FPKM). d Top 10 biological functions of upregulated genes after R-spondin2 treatment using GO analysis. e GSEA comparison showing upregulation of mesoderm-associated genes with R-spondin2 treatment. f The kinetics of expression of representative mesoderm genes BRACHYURY, MIXL1, APLNR, and KDR during early hematopoietic differentiation from hESCs in chemically defined condition with or without treatment of R-spondin2 (20 ng/mL) analyzed with real-time PCR. Relative expression is normalized to the level (= 1) of control at day 0. Results are shown as means ± SD (n = 3). Results are shown as means ± SD (n = 3). *P < 0.05, ***P < 0.001