Antibody against ki 67

The tumor tissues were made into paraffin sections as previously described. The sections reacted with antigen retrieval buffer in boiling for 10 min, followed by blocking with 3% H₂O₂ for 10 min and 1% BSA for 15 min. Subsequently, the sections were incubated with antibody against Ki-67 (1:50; Affinity) at 4°C overnight, and HRP-labeled secondary antibody (1:500; Thermo Fisher Scientific Co., Ltd.) at 37°C for 60 min. The sections then reacted with DAB reagents for several minutes and were counterstained with hematoxylin. Finally, the sections were dehydrated with ethanol and xylene, mounted with gum, and photographed with a microscope at 400× magnification.

Quercetin (purity ≥98.5%) and Temozolomide (TMZ) (purity ≥98%) were purchased from Aladdin (Shanghai, China) and dissolved in dimethyl sulfoxide (DMSO). Cell Counting Kit-8 was purchased from Bimake (Houston, TX, United States). Antibodies against GSK-3β, β-catenin, E-cadherin, N-cadherin, Vimentin, ZEB1, Akt, p-Akt and anti-rabbit IgG (Alexa Fluor^® 488 Conjugate) were purchased from Cell Signaling Technology (Danvers, MA, United States). Antibodies against β-actin, p-GSK-3β (Ser 9) were purchased from Santa Cruz (Dallas, Texas, United States). Horseradish peroxidase-conjugated goat anti-rabbit IgG, Horseradish peroxidase-conjugated goat anti-mouse IgG secondary antibodies and Phalloidin were purchased from Zhongshan Jinqiao Biotechnology Co. Ltd. (Beijing, China). Antibody against Ki67 was obtained from Affinity Biosciences (Zhenjiang, China). The SDS-PAGE Gel Quick Preparation Kit, RIPA lysis buffer, Crystal violet and BCA Protein Assay Kit were purchased from Beyotime Biotechnology (Shanghai, China). SB216763 was purchased from MedChemExpress (New Jersey, United States).