Hematoxylin staining solution

FA aqueous solution (37% w/v) was from Sigma Aldrich (USA); methanol solution (99.5%) was from Xilong Scientific Co., Ltd. (Shenzhen, China). Mangiferin (MGF) standards (≥98%) were obtained from Dibai Biotechnology Co., Ltd. (Shanghai, China); 4-phenylbutyric acid (4-PBA) was from Aladdin (Shanghai, China); SB216763 and KN-93 phosphate standards were from Yuanye Bio-Technology (Shanghai, China); Dulbecco’s modified Eagle’s medium (DMEM)/F12, phosphate buffer solution, penicillin streptomycin mixture (100×), trypsin solution (0.25%), and special grade fetal bovine serum (FBS) were products from Biological Industries (Israel); dimethyl sulfoxide (DMSO, cell culture grade) was obtained from Solarbio (Beijing, China); Cell Counting Kit-8 (CCK8) came from Proteintech (Wuhan, China); BCA protein assay kit, RIPA lysate, DCFH-DA (ROS detection probe), and DAPI staining solution were from Share-bio (Shanghai, China); and calcium content chromogenic assay kit and Mito-Tracker Red CMXRos (mitochondrial red fluorescent probe) were obtained from Beyotime Biotechnology (Shanghai, China). Citric acid antigen repair solution, hematoxylin staining solution, and DAB color reagent were from Servicebio technology Co., Ltd. (Wuhan, China). Other analytical grade reagents, unless otherwise specified, were obtained from a local reagent vendor.

The tissue microarray (TMA) of odontogenic lesions including 11 cases of DC and 30 cases OKC was constructed by Servicebio, Wuhan, China. The TMA of odontogenic lesions was incubated in 65°C for 2 h. The slides of TMA were dewaxed by xylene and dehydrated by graded ethanol. For antigen exposure, the slides were boiled in the electric pressure cooker and cooked to 120°C for 60 seconds. After cooling to the room temperature, the slides were blocked by goat serum. The primary antibody IL-8 (1 : 200, Proteintech, China) was incubated with the slides overnight and followed by HRP-conjugated solutions. Finally, the slides were stained with diaminobenzidine (Zhongshan Golden Bridge Biotechnology Co. Ltd., China) and hematoxylin staining solution (Servicebio, Wuhan, China).