Anti hemagglutinin ha

The cell samples or frontal lobes of brain tissues were lysed with radioimmunoprecipitation assay buffer (Thermo Scientific, 87788), and 20 μg of samples was separated with 4 to 20% SDS–polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane (Millipore, Shanghai, China). After blocking, the membrane was incubated overnight with the indicated primary antibody at 4°C. The following primary antibodies were used: anti-GFAP (1:10,000; Abcam, ab7260), anti-NDRG2 (1:2000; Abcam, ab174850), anti-p-Smad2 (Ser^465/467)/Smad3 (Ser^423/425) (1:1000; Cell Signaling Technology, 8828), anti-Smad2/3 (1:1000; Cell Signaling Technology, 8685), anti-Smad3 (1:1000; Cell Signaling Technology, 9528), anti–MMP-9 (1:1000; Santa Cruz Biotechnology, sc21733), anti–laminin α2 (1:200; Sigma-Aldrich, L0663), anti-occludin (1:10000; Proteintech, 66378-1), anti–ZO-1 (1:1000; Proteintech, 66452-1), anti-flag (1:5000; Sigma-Aldrich, F1804), anti-hemagglutinin (HA; 1:20000; Proteintech, 66006-2), and anti–β-actin (1:5000; Abcam, ab8226). Horseradish peroxidase–conjugated secondary antibodies were used. Chemical reactions were detected with an ECL system (Advansta, Menlo Park, CA, USA). The scanned images were analyzed with ImageJ NIH software.

The following primary antibodies were used: anti-ZMYND8 (Proteintech), anti-ACTB (Actin Beta) (Proteintech), anti-hemagglutinin (HA; Proteintech), anti-Flag (Proteintech), anti-ubiquitin (Proteintech), anti-H3K4me3 (Cell Signaling Technology), anti-KDM5C (Bethyl), anti-ZNF592 (Bethyl), and anti-ZNF687 (Bethyl). The HRP-linked anti-mouse (1/5000) and anti-rabbit antibodies (1/5000) were purchased from Cell Signaling Technology.