Immunohistochemistry of miR-143 in Oral Tumors

Human oral tumor IHC was performed according to previous descriptions [16 (link)]. Briefly, sections of miR-143 low or high expression were deparaffinized in xylene. Then, samples were rehydrated through a gradient concentration of alcohol followed by incubation by 5% normal goat serum to block non-specific staining. Sections were then incubated with rabbit anti-HK2 antibody (1:200, Cellsignaling, #2867, Danvers, MA, U.S.A.) overnight at 4°C. The slides were incubated with biotin-labeled goat anti-rabbit IgG and further incubated with streptavidin peroxidase solution (SABC kit, Boster Biological Technology, Ltd., Wuhan, China). The staining was visualized by reaction with 3,3′-di-aminobenzidine (Boster Biological Technology, Ltd.) in PBS for 5 min at room temperature. All of the IHC staining results were reviewed independently by two pathologists.