Fig. 4

The role of RSPO1 on eMSC clonogenicity and phenotypic expression. a Gene expression of RSPO1 in stromal cells from the menstrual, proliferative and secretory phase using qPCR (n = 7). b Protein expression of RSPO1 in stromal cells from the menstrual, proliferative and secretory phase using western blotting. (n = 7). c Representative immunofluorescent images showing unfractionated endometrial stromal cells and eMSCs (CD140b⁺CD146⁺cells) expressing RSPO1 (red). d Relative cloning efficiency of eMSC colonies in growth medium, epithelial cell CCM from the menstrual phase, epithelial cell CCM with addition of rabbit IgG and epithelial cell CCM with addition of RSPO1 antibody at 1 μg/ml (n = 7). e Relative proportion of CD140b⁺CD146⁺ cells after treatment (n = 7). Data were normalized to control. f The TCF/LEF luciferase signal of eMSCs with epithelial CCM from the menstrual phase; epithelial cell CCM with addition of rabbit IgG and epithelial CCM with addition of human anti-RSPO1 antibody at 1 μg/ml. Data normalized to growth medium (n = 5). g Relative cloning efficiency of eMSC colonies in growth medium, 50 ng/ml of rhRSPO1, 50 ng/ml of rhRSPO1 + 25 ng/ml of rhWNT3A and 25 ng/ml of rhWNT3A alone (n = 7). h Relative proportion of CD140b⁺CD146⁺ cells after treatment (n = 7). Results are shown as mean ± SEM; *P < 0.05, **P < 0.01, *** P < 0.001. Scale bar, 50 μM. CCM, concentrated conditioned medium; eMSCs, endometrial mesenchymal stem-like cells; rh, recombinant human