Gb11264

The slices were stained with hematoxylin and eosin (H&E), Masson’s trichrome staining, and immunohistochemical (IHC) staining which incubated with primary antibodies against Runx2 (GB11264, ServiceBio, China), Col I (GB11022-3, ServiceBio, China), and Ocn (GB11233, ServiceBio, China), respectively. Images were taken using a Leica Biosystems microscope (Aperio^® AT2, Germany).

The mandible samples were decalcified in a fast decalcified solution (Biotech, China) for 24 h at room temperature, dehydrated in graded ethanol and then embedded in wax. The tissue was cut into 5 μm sections using a microtome (Leica) and stained with HE, Masson’s trichrome and TRAP with commercial staining kits (Servicebio) to assess new bone formation and osteoclast function. After being blocked with bovine serum albumin (Servicebio), the sections were incubated with primary antibodies against Arg-1 (1:500, GB11285, Servicebio) and Runx2 (1:500, GB11264, Servicebio) as well as CD86 (1:500, 26903-1-AP, Proteintech) and Osx (1:500, ab209484, Abcam) at 4 °C overnight. Then, the sections were incubated with appropriate secondary antibodies (Servicebio) before mounting with DAPI. Samples were imaged using a DMI8 inverted fluorescence microscope (Leica Microsystems, Germany) and an LSM980 laser scanning confocal microscope with quantitative analysis by ImageJ.