Neutral gum

Dewaxed sections were incubated with hematoxylin (Servicebio) and eosin (Servicebio); dehydrated with xylene, anhydrous alcohol, and 75% ethanol; and sealed with neutral gum (Servicebio). The cytoplasm and nucleus of all the cells turned to red and blue, respectively. The slices were scanned (Aperio AT2, Leica), and osteoclasts and tooth resorption lacuna were observed at the pressure side of the upper-left first molars.

All tissue samples were fixed in 4% paraformaldehyde solution for 12 h, and tissue samples were embedded using paraffin wax. Hematoxylin kit (Servicebio) and Eosin staining kit (Servicebio) were used for staining, followed by sealing with neutral gum (Servicebio). For Ki67 staining, closure was performed using 5% bovine serum albumin (Servicebio). The samples were then incubated with anti-Ki67 polyclonal antibody (1:800 dilution) (Servicebio) and fluorescently labeled secondary antibody (1:2000 dilution) (Zhuangzhi, Shaanxi, China), and captured by fluorescence microscopy. Immunohistochemical staining was performed using tumor tissue sections from TNBC patients. The sections were permeabilized and blocked with 3% hydrogen peroxide, and then incubated with CCDC150 primary antibody (CUSABIO, Wuhan, China). The images were captured by fluorescence microscopy.