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Fig. 3 | Stem Cell Research & Therapy

Fig. 3

From: Type I collagen deposition via osteoinduction ameliorates YAP/TAZ activity in 3D floating culture clumps of mesenchymal stem cell/extracellular matrix complexes

Fig. 3

Osteoinductive medium facilitates F-actin formation and YAP/TAZ activity in C-MSCs. a, b C-MSCs were cultured in OIM for the indicated culture period. a Confocal immunofluorescence images of COL1 and nuclei. Bar = 100 μm. b Confocal immunofluorescence images of YAP/TAZ (green), F-actin (red), and nuclei (blue) in C-MSCs. Bar = 20 μm. White boxes show enlarged images. c The graph summarizes the distribution of YAP/TAZ localization patterns. The patterns were classified as mainly nuclear (N > C), diffuse (N = C), or mainly cytoplasmic or undetectable (N < C or undetectable). df C-MSCs: C-MSCs cultured in GM for 5 days. OIM-C-MSCs: C-MSCs cultured in OIM for 5 days. d Immunoblotting for YAP/TAZ. A rabbit anti-YAP/TAZ (D24E4) mAb (Cell Signaling) was used to detect both the YAP and TAZ proteins (middle panel). To show YAP expression more clearly, we also used a rabbit anti-YAP (D8H1X) mAb (upper panel). e Real-time PCR of YAP/TAZ target genes. Data were normalized to the values of C-MSCs. Values represent means ± S.D. of three cultures (**p < 0.01). f C-MSCs transfected with negative control or YAP and TAZ siRNAs were cultured in OIM for 5 days. RUNX2 expression levels were analyzed by real-time PCR. Data were normalized to the values of OIM-C-MSCs transfected with negative control siRNA. Values represent means ± S.D. of three cultures (**p < 0.01). All graphs and images are representative of three independent experiments

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