N cor

The brain tissue and cell sections were prepared. After permeabilization with 0.3% Triton X-100 (Sigma-Aldrich), sections were probed with LAMP-2A (1:100, Abcam, ab18528), N-CoR (1:100, Novus, NBP1-28863) for tissues or N-CoR (1:100, Santa Cruz, sc-1609) for cells. After being washed with PBS containing 0.05% of Tween-20 (Sigma-Aldrich), sections were incubated with fluorescent secondary antibodies (Beyotime, China) and antifade mounting medium containing 1:500 DAPI (Beyotime, China). Leica fluorescent microscope was used for visualization.

Immunohistochemistry was performed with LAMP-2A (1:100, Abcam, ab18528), N-CoR (1:100, Santa Cruz, sc-1609) for cells and N-CoR (1:100, Novus, CO, US, NBP1-28,863) for tissues. After being equilibrated at − 20 °C for 15 min, brain tumor tissues were sectioned at a range of 5–7 μm and fixed with 4 °C pre-cold acetone. Cultured cells growing on glass slides were fixed with 4% of formaldehyde. The tissue or cell sections were incubated with 3% of H₂O₂ for 10 min, followed by 10% of BSA as blocking solution for 1 h at room temperature. Then the sections were incubated with the primary antibodies overnight at 4 °C and hybridized with horseradish peroxidase labeled secondary antibody (Beyotime, China) for 1 h at room temperature. The diaminobenzidine kit (DAB kit; Long Island Biotec, Shanghai, China) was applied for visualization and hematoxylin (BASO, China) for counterstain.