Fig. 2

Macrophages promote the proliferation and migration of ESCs via the CCL20/CCR6 axis in EMs. A ELISA analysis of CCL20 in ESC culture supernatant with or without co-culture treatment. B The mRNA level of CCL20 in ESCs with or without co-culture treatment by qRT-PCR. C The protein level of CCR6 in ESCs in different groups by western blot. D Quantification of the gray value of western blot bands. E Representative immunofluorescence images of CCR6 (red) in ESCs in different groups, Nuclei were stained with DAPI (blue). (original magnification 200 ×) F Quantitative fluorescence intensity of CCR6. G Proliferative capability of ESCs in different groups assessed by CCK8 assay. H Proliferative capability of ESCs in different groups assessed by Edu assay. (original magnification 200 ×) I Quantification of Edu incorporation, Edu incorporation rate (%) = Edu positive cells (green)/Hoechst positive cells (blue). J Migration capability of ESCs in different groups assessed by wound healing assay. (original magnification 40 ×) K Quantification of wound closure rate, wound closure rate (%) = (Scratch distance 0 h—Scratch distance 24 h) / Scratch distance 0 h. L Migration capability of ESCs in different groups assessed by Transwell assay. (original magnification 200 ×) M Quantification of migrating cells per field at 200 × magnification. Data are presented as the mean ± SD of n = 3 independent experiments. Statistical analysis was performed using Mann–Whitney test (A, B) or one-way ANOVA with Tukey post hoc (D, F, G, I, K, M). ****p < 0.0001, ***p < 0.001, **p < 0.01, ns, non-significant vs. control group. ^####p < 0.0001, ^####p < 0.001, ^##p < 0.01 vs. co-culture group