Fig. 3
TGF-β-Smad3 signaling induces cell cycle arrest in the colonic epithelial organoids of old mice. a Cell cycle analysis through PI staining using flow cytometry (left) and quantitative measurement of the cell cycle phase in the young- and old-mouse organoids (rignt). b Relative levels of elF4E mRNA expression in colonic epithelial organoids from young and old mice. c Protein expression levels of cyclin D1 and β-actin in young- and old-mouse organoids. The images were cropped from the original blot image in Supplementary Fig. 6a. The blot was cut before hybridization with antibodies. d mRNA expression of cell cycle-regulated genes in young and old-mouse organoids. Data are shown as ± standard error of the mean (SEM) (n = 7). e ID1, ID2, and ID3 mRNA expression in young- and old-mouse organoids. Data are reported as the mean ± SEM (n = 7). f Protein expression levels of ID1, ID2, ID3, and β-actin in young- and old-mouse organoids. The images were cropped from the original blot image in Supplementary Fig. 6b. The blot was cut before hybridization with antibodies. g Protein expression levels of p-16INK4a and β-actin in young- and old-mouse organoids. The images were cropped from the original blot image in Supplementary Fig. 6c. The blot was cut before hybridization with antibodies. *p < 0.05, **p < 0.01, ***p < 0.001,.****p < 0.0001 by the Mann–Whitney t-test. TGF-β, transforming growth factor β; ID1, inhitor of DNA binding 1; ID2, inhitor of DNA binding 2; ID3, inhitor of DNA binding 3