Fig. 3
Detection of tumor necrosis factor-α (TNF-α)-induced osteocyte necroptosis in the mouse model of orthodontic tooth movement (OTM). a Hematoxylin and eosin (H&E) staining of the region of interest (ROI) on the compression side in wild-type (WT) mice and TNF receptor type 1- and 2-deficient (TNFRsKO) mice. Scale bar = 50 μm. b Percentage of dying osteocytes (orange) and empty lacunae (red) in WT and TNFRsKO mice (n = 6). c Immunofluorescence staining for phosphorylated (p)-RIP3 (red) and DAPI staining (blue) of ROIs on the compression and tension sides in WT and TNFRsKO mice. Scale bar = 50 μm. d High-magnification images of the compression side on day 6 after OTM in WT mice. White arrows indicate p-RIP3-positive osteocytes. Scale bar = 50 μm. e Percentage of p-RIP3-positive osteocytes on the compression and tension sides of WT and TNFRsKO mice (n = 6). f Immunofluorescence staining for p-MLKL (red) and DAPI staining (blue) of ROIs on the compression and tension sides in WT and TNFRsKO mice. Scale bar = 50 μm. g High-magnification images of the compression side on day 6 after OTM in WT mice. White arrows indicate p-MLKL-positive osteocytes. Scale bar = 50 μm. h Percentage of p-MLKL-positive osteocytes on the compression and tension sides in WT and TNFRsKO mice (n = 6). The dotted line indicates the border between the alveolar bone and periodontal ligament. a, alveolar bone; p, periodontal ligament. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
