Fig. 1

R-spondin2 promotes generation of hematopoietic progenitors from hESCs. a Left panel: heatmap of Rspo expression in OP9-d4, OP9-d8, and MS5 stromal cells (accession number GEO: GSE61580). Right panel: heatmap of Rspo expression in AGM-S3-A7, AGM-S3-A9 subclones of AGM-S3 stromal cell and OP9 cells (accession number GEO: GSE11891). b Real-time PCR analysis of expression of Rspos in mAGM-S3 stromal cells. Relative expression is normalized to the level (= 1) of Actin. Results are shown as means ± SD (n = 3). c Representative immunofluorescence images of H1 cells with or without treatment of R-spondin2 (20 ng/mL) showing the generation of CD43⁺ HPCs at day 7 of mAGM-S3 co-culture. d Flow cytometry analysis of H1 cells with or without treatment of R-spondin2 (20 ng/mL) showing the generation of CD43⁺ HPCs at day 7 of mAGM-S3 co-culture. Results are shown as means ± SD (n = 3). ***P < 0.001. e Representative immunofluorescence images of H1 cells with or without the treatment of R-spondin2 (20 ng/mL) showing the generation of CD45⁺ HPCs at day 10 of mAGM-S3 co-culture. f Flow cytometry analysis of H1 cells with or without the treatment of R-spondin2 (20 ng/mL) showing the generation of CD45⁺ HPCs at day 10 of mAGM-S3 co-culture. Results are shown as means ± SD (n = 3). **P < 0.01. g Hematopoietic colony-forming potential of hESC-derived cells after 12 days of co-culture with or without the treatment of R-spondin2 (20 ng/mL), analyzed using total colony numbers (upper panel) and the proportion of BFU-E (burst-forming unit-erythroid), CFU-E (colony forming unit-erythrocyte), CFU-GM (colony forming unit-granulocyte/macrophage) and CFU-GEMM (colony-forming unit-granulocyte/erythroid/macrophage/monocyte) (lower panel). Results are shown as means ± SD (n = 3). NS, not significant, *P < 0.05. GEMM, P = 0.56; GM, P = 0.32; CFU-E, P = 0.65; BFU-E, P = 0.64