Immunohistochemistry of Ameloblastoma

The ameloblastoma samples were obtained from Guanghua School of Stomatology, Sun Yat-sen University. The tumors were fixed in 4% paraformaldehyde for 24 h, dehydrated, embedded in paraffin, and sectioned at 4 μm thickness. After dewaxing, the sections were subjected to heat-induced epitope recovery (Zsbio, Cat#ZLI-9065) and incubated in 3% H₂O₂ for 10 minutes. Then, the sections were washed and incubated with primary antibodies against KI67 (Novus, Cat#NB500-170; 1:200) and EZH2 (Cell Signaling Technology, Cat#5246 S; 1:200) overnight at 4 °C. After incubation with the secondary biotinylated antibody (Zsbio, Cat#PV-6001-18) for 40 minutes, horseradish peroxidase conjugation was carried out with a DAB kit (Zsbio, Cat#ZLI-9017). The scores of each sample were assessed via the staining intensity and area of tumor cells as described previously.^{25 (link)} For the H&E assay, paraffin-embedded tumor sections were stained with an H&E kit (Solarbio, Cat#G1120-100) following the manufacturer’s instructions.