Fig. 1

YAP and TAZ are downregulated in 3D-cultured C-MSCs. a Schematic image of C-MSC culture. Bar = 5 mm. b–g C-MSCs were generated and maintained in growth medium (GM) for the indicated culture periods. b Upper panels indicate macroscopic images of C-MSCs. Lower panels show confocal immunofluorescence images of COL1 and nuclei in C-MSCs. Bar = 100 μm. c Confocal immunofluorescence images of YAP/TAZ (green), F-actin (red), and nuclei (blue) in C-MSCs. Bar = 20 μm. White boxes show enlarged images. d The graph summarizes the distribution of YAP/TAZ patterns. The patterns were classified as mainly nuclear (N > C), diffuse (N = C) and mainly cytoplasmic or undetectable (N < C or undetectable). e Immunoblotting for YAP/TAZ in C-MSCs. f, h Real-time PCR for YAP and TAZ (f) or YAP/TAZ target genes (h) in C-MSCs. Data were normalized to the values on day 0. Values represent means ± S.D. of three cultures (**p < 0.01). g Phos-tag-SDS-PAGE was used to show the phosphorylation levels of YAP/TAZ. Arrowheads indicate the position of unphosphorylated YAP/TAZ. Levels of total YAP/TAZ were analyzed by immunoblotting. All graphs and images are representative of three independent experiments