Fig. 2: IL-33/ST2 increased the survival rate and migration ability of eESCs.

A The eESCs were treated with specified concentrations of human recombinant IL-33 protein (rIL-33) (25, 50, 100, and 200 ng/ml) for different times (12, 24, 48, and 72 h). CCK-8 assays were performed to detect cell viability in different groups. B RT-qPCR was used to determine the relative levels of ST2 mRNA in eESCs transfected with siST2 (50 nM), sicon (50 nM) for 48 h. C Western blot was used to determine the protein levels of ST2 in eESCs transfected with siST2 (50 nM) or sicon (50 nM) for 48 h. D Transwell migration assay was performed to detect the migrant ability of eESCs treated with or without macrophages co-culture. Cartoon picture showed the experimental progress. (original magnification ×200). E, F Transwell migrantion assays were performed in rIL-33 (100 ng/ml) treated eESCs (E) and siST2 (50 nM) transfected eESCs (F). (original magnification ×200). G The wound healing assays were conducted in eESCs transfected with siST2 (50 nM) and control group. (original magnification ×40). H Colony formation assays were performed in siST2 (50 nM) transfected eESCs treated with rIL-33 (100 ng/ml) or macrophages co-culture. (original magnification ×40).Data are presented as the mean ± SD, n = 3 independent experiments. Statistical analysis was performed using Student’s t test (B–D, G) or 2-way ANOVA (H). ****p < 0.0001, ***p < 0.001, *p < 0.05, ns, non-significant. sicon negative control siRNA, siST2 siRNA targeting ST2, M macrophages co-culture treatment.