Fig. 2

WNT16 treatment inhibited mineralization and promoted cell senescence of AS-osteoprogenitor cells during osteoblast differentiation. Both control and AS-osteoprogenitor cells were differentiated with osteogenic medium in the presence of vehicle or WNT16. Analysis of A ALP staining, B ARS staining, and VON staining, C HA staining, D intercellular ALP activity at indicated day, E quantitation of ARS and VON at day 21, and HA staining at day 28 (n=4 per each group). F SaOS2 cells were co-transfected with 3 μg ALP, BSP, OSE, or OCN promoter plasmid along with β-gal plasmid for 48 h followed by and WNT16 treatment for 24 h and then analyzed by luciferase assay (n=6 per each group). G SA-β-gal staining was performed at 28 days of osteoblast differentiation of control and AS-osteoprogenitor cells. Counting results of Fig. 3G (right panel) (n=4 per each group). AS-osteoprogenitor cells were differentiated into mature osteoblasts in the presence of vehicle and WNT16 (50ng/ml). Analysis of H immunoblotting for protein level and I RT-qPCR for mRNA level (n=3 per each group). Scale bar = 200 μm. Data are presented as the median and interquartile range. *p<0.05