Fig. 2: Piezo1 is required for osteocyte maturation and dendrite formation.

a Phalloidin staining (green; nucleus: blue) and osteocyte dendrite quantification of femur midshaft cortical bone in 8-week-old WT and Dmp1Cre; Piezo1^f/f (Piezo1 cKO) mice. N = 3 biological replicates. b Analysis and quantification of osteocyte dendrite orientation in 8-week-old WT and Piezo1 cKO mice. Color bar indicates the fiber angle with respect to the long bone axis. Axial (red): 0°−30°. Oblique (yellow): 30−60°. Transverse (cyan): 60−90°. N = 3 biological replicates. c Silver nitrate staining and quantification of LCN in 8-week-old WT and Piezo1 cKO mice. N = 3 biological replicates. d IF staining and quantification of SOST expression (cyan; nucleus: blue) in 8-week-old WT and Piezo1 cKO mice. N = 5 biological replicates. e IF staining and quantification of SPP1 expressions (red; nucleus: blue) in 8-week-old WT and Piezo1 cKO mice. N = 4 biological replicates. f QRT-PCR analysis of mRNA expressions of osteocyte (Sost, Dmp1, Phex) and osteoblast genes (Col1A1, Sp7, Spp1) in cortical bone tissue of 8-week-old WT and Piezo1 cKO mice. N = 5 biological replicates. g Analysis of embedding osteocytes (red) in femur midshaft cortical bone from Sox9-CreER^T2; TdTomato and Sox9-CreER^T2; Piezo1^f/f; TdTomato mice. Red: tdTomato. Blue: nucleus. TAM was injected at P21. Mice were collected 5-weeks post-TAM injection. N = 3 biological replicates. h Phalloidin staining (green; tdTomato: red) of femur midshaft cortical bone from Sox9-CreER^T2; TdTomato and Sox9-CreER^T2; Piezo1^f/f; TdTomato mice, quantified in Supplementary Fig. 2k. N = 5 biological replicates. Scale bars: (a–e, g, h) 50 µm. Bar graphs represent mean ± SD. (a–g) Two-sided unpaired Student’s t test. Source data are provided as a Source Data file.