Fig. 6

The expression levels of Wnt signaling pathway-related genes and proteins were decreased in Epi-GMSCs. (A) RT-qPCR analysis of the expression of LRP5, LRP6, COL1A1, β-catenin, RUNX2, WNT3, WNT5A, WNT11, FZD5, FZD7, and TCF7L2 after days 3, 9, and 15 of incubation in GECCM (n = 3, ****P < 0.0001). (B) WB and (C) quantitative analysis of the expression of LRP5, COL1A1, β-catenin, and RUNX2 after days 3, 9, and 15 of incubation in GECCM. Full-length blots/gels are presented in Supplementary Fig. 4. (n = 3, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). (D) Quantitative analysis of β-catenin immunofluorescence intensity using ImageJ. (n = 3, *P < 0.05). (E) Representative confocal fluorescence images showing the subcellular localization of β-catenin (red). Cytoskeleton analysis was performed by immunofluorescence staining with F-actin (green). (n = 3)