Osteocyte mTORC1 in mice regulates granulopoiesis. (A) The level of P-S6 (S235/236) in primary osteocytes isolated from 12-week-old Dmp1-TSC1, Dmp1-Rheb, and control mice was detected by western blotting. (B) The TSC1 and Rheb mRNA expression in osteocytes, heart, liver, kidney, spleen, and lung tissues from 12-week-old Dmp1-TSC1, Dmp1-Rheb, and control mice was detected by qPCR. (C) Immunofluorescence microscopy of femur sections of 12-week-old Dmp1-TSC1, Dmp1-Rheb, and control mice. Cells were stained with anti-Dmp1 (red) and anti–P-S6 (green). Blue, nuclei visualized by 4′,6-diamidino-2-phenylindole (DAPI) stain. Scale bar, 50 µm. (D) The total number of PB neutrophils in 12-week-old Dmp1-TSC1, Dmp1-Rheb, and control mice (mean ± standard deviation [SD]; n = 10). (E) FACS analysis of BM and PB CD11b⁺Gr-1⁺ neutrophils in 12-week-old Dmp1-TSC1, Dmp1-Rheb, and control mice (mean ± SD; n = 10). Data are mean ± SD of 3 independent experiments. **P < .01; ***P < .001.