Fig. 3

The nuclear-cytoplasmic shuttling of YAP in MSCs after co-cultured with fibroblasts. A A total of 1 × 10⁴ MSCs were cultured either alone or with 3 × 10⁴ fibroblasts in a single well of a 12-well plate. The cells were treated either with a vehicle or with 200 ng/mL BMP2 for 2 h. Subsequently, the cells were harvested and subjected to immunofluorescence. Representative immunofluorescent staining images were shown. The colour green was used to represent YAP, while red indicated α-SMA and blue represented DAPI. Scale bars, 100 μm for ×400, 20 μm for ×630. M MSC, F fibroblast. B 3 × 10⁴ fibroblasts were cultured on the upper layer of Corning cell culture insert with 0.4 μm polyester membrane. 1 × 10⁴ MSCs were placed below the cell permeable membrane for 2 h, supplemented either with a vehicle or with 200 ng/mL BMP2. Subsequently, the cells were harvested and subjected to immunofluorescence. Representative immunofluorescent staining images of MSCs were presented. Green colour represented YAP, while blue indicated DAPI. Scale bars, 25 μm for ×400, 10 μm for ×630. C, D Quantitative analysis of nuclear/cytoplasmic YAP intensity of MSCs in A and B was shown. The average nuclear/cytoplasmic YAP intensity of at least ten cells were calculated and presented (*P < 0.05; **P < 0.01). E 3 × 10⁴ fibroblasts were cultured on the upper layer of Corning cell culture insert with 0.4 μm polyester membrane. 1 × 10⁴ MSCs were placed below the cell permeable membrane for the indicated time point, supplemented either with a vehicle or with 200 ng/mL BMP2. Subsequently, the cells were harvested and subjected to immunoblotting. Images of western blots to show the expression of YAP and p-YAP in MSCs were presented. Relative band intensity was shown below the gels and full-length gels were presented in Additional file 1: Fig. S7A