Fig 2: LiCl promoted osteogenesis in OVX BMSCs by regulating of autophagy. a, Real-time qPCR analyses of Beclin-1, Atg8/LC3, and p62 mRNA expression in different groups. b, Real-time qPCR analyses of Runx2, Alp, Opn, Opg, and Rankl expression in different groups. c, Western blotting analysis of BECLIN-1, LC3II, p62, ALP, OSX and RUNX2 in different groups. d, Quantification analysis of the expression of BECLIN-1, LC3II and p62 protein. e, Quantification analysis of the expression of ALP, OSX, and RUNX2 protein. Data represent means ± S. D (n = 3).

Fig 3: Inhibition of Piezo1 suppressed osteoclastic activities on the tension side. TRAP staining (A) and quantification of TRAP-positive cells (D). TRAP-positive osteoclasts were detected near the tension side of the roots. On day 3, the number of osteoclasts in the OTM group was significantly greater than that of the CON and the OTM + INH groups. Scale bar = 50 μm. Immunohistochemical staining of RANKL (B) and OPG (C) of PDL on the tension side of the distobuccal roots. The means of the integrated optical density (IOD) of RANKL (E), OPG (F), and RANKL/OPG ratio (G) were shown. On day 3, the RANKL/OPG ratio of the OTM group was significantly greater than that of the CON and OTM + INH groups. Western blot analyses (H) were carried out to detect the protein levels of RANKL and OPG on day 3 of OTM. The protein expression of the RANKL/OPG ratio increased significantly in the OTM group and was reduced by GsMTx4 in the OTM + INH group. Data are presented as the mean ± SD. *p < 0.05. Scale bar = 50 μm. TRAP, tartrate-resistant acid phosphatase; RANKL, receptor activator of nuclear factor-kappa B ligand; OPG, osteoprotegerin; AB, alveolar bone; R: root.

Fig 4: LiCl promoted autophagy, inhibited apoptosis and osteoclastogenesis during OTM in mice. a, Immunohistochemical staining of Beclin-1 in the tension area of OTM in different groups on day 7 and day 14. b, Immunohistochemical staining of OPG and RANKL in the tension area during OTM in different groups on day 14. c, TUNEL examination of cell apoptosis in the tension side area on day 3. Green fluorescence represents apoptotic cells. d, The MOD of Beclin-1 in the tension area on day 7 and day 14. e, The ratio of OPG/RANKL in the tension area during OTM on day 14. f, Quantification analysis of the apoptotic cells. Data represent means ± S. D (n = 3). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)