Fig. 3

WNT16 treatment facilitated cell senescence of AS-osteoprogenitor cells under H₂O₂-stimulation. A Experimental design for cell senescence treated with WNT16. AS-osteoprogenitor cells were exposed to H₂O₂ for 2 h and then incubated with vehicle or WNT16 treatment for 70 h (n=5 per each group). Analysis of B SA-β-gal staining, C counting results of SA-β-gal-positive cells, D assessment of osteoprogenitor cell lysates using hydrogen peroxidase assay (n=5 per each group), E immunoblotting for protein level, F protein quantification of Fig. 3E, and G RT-qPCR for mRNA level (n=4 per each group). H SaOS2 cells were transfected with TOP or FOP-flash in the presence β-galactosidase for 24 h, incubated with WNT16 treatment for 24 h, and then analyzed by luciferase assays (n=4 per each group). I Experimental design for cell senescence treated with siRNA against WNT16. AS-osteoprogenitor cells were transfected with 100 nM WNT16 siRNA #2 for 70 h, exposed to H₂O₂ for 2 h, and followed by analysis of H immunoblotting for WNT16 protein level and I qPCR for mRNA level (n=5 per each group). Data are presented as the median and interquartile range. *p<0.05, **p<0.01. Representative images are shown. Scale bar = 200 μm