Fig. 6

CM and E2-CM promoted angiogenesis in vitro and in vivo, possibly with angiogenin involvement. A Effects of CM/E2-CM on restoring CTX-affected ovarian angiogenesis in mice. Confocal images of IF staining (a) and quantitative analysis (n = 5) (b) for angiogenic marker CD31 (in green) and VEGF-A (in red) in ovary tissues of each experimental group. Immunoblotting results for CD31 and VEGF-A (c) and their and quantitative analysis (n = 3) (d). B Protein component analysis of CM and E2-CM. Results obtained through cytokine array analysis. Nonconcentrated medium pool from pcMSC passages 8–11. Total protein, 700 µg/ml. (a). Top eight abundant cytokines (b).). The concentration of Angiogenin in CM and E2-CM was shown. ELISA, **p < 0.01, Student t test (n = 8) (c). C Effects of CM and E2-CM on angiogenesis, as revealed through tube formation assay using human HUVEC cells. Fluorescent images depicting tube formation labeled with Calcein AM in experimental groups. Anti-angiogenin antibody used to neutralize angiogenin (a). Quantitation and statistical analysis of total tube length (b) and branching point number (c) under various experimental condition (n = 5). *p < 0.05, **p < 0.01, ***p < 0.001. One-way ANOVA with Tukey’s test