Fig. 3

Comparative study of DETCs and EpSCs with mixed and Transwell coculture systems. A–B EpSCs were isolated from neonatal WT mice and cultured to passage 2–4 before analysis. EpSCs were cocultured with DETCs in a mixed coculture system (DETCs group) or noncontact Transwell coculture system (DETCs-Transwell group). The proportions of CD49f^briCD71^dim cells (A) and K15⁺ cells (B) were detected by FACS after 3 days of coculture. C EpSCs were isolated from neonatal WT mice and cultured for three days before labeling with CFSE. CFSE-labeled EpSCs were cocultured with DETCs in a mixed coculture system (DETCs group) or noncontact Transwell coculture system (DETCs-Transwell group). The proportion of CFSE^low cells was detected by FACS after 3 days of coculture. D–F DETCs-derived exosomes were isolated from the culture medium of DETCs. The particle size distribution of DETCs-derived exosomes was measured using a nanoparticle sizer (D). The expression of typical proteins (Calnexin, CD63, β-actin and TSG101) in DETCs-derived exosomes and the control group (cellular protein from 3T3 cells) was detected by means of WB (E). The morphology of DETCs-derived exosomes was detected by transmission electron microscopy (Black arrows indicate Exo) (F). All data are representative of at least three independent experiments and represent mean ± SD of indicated number of mice per group. The p value was calculated by one-way ANOVA with Bonferroni’s multiple comparison test (A–C) (^nsp > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001)