A19626

Protein samples from cells and tissues were extracted and total protein concentration was determined using a BCA kit (Beyoncé, P0012, Shanghai, China). The total protein in the samples was separated by 6%–12% SDS-polyacrylamide gel electrophoresis and then transferred to 0.22 μm polyvinylidene difluoride membranes. Subsequently, the membranes were incubated with primary antibodies, followed by incubation with the corresponding secondary antibodies. The target bands were developed using the ECL chemiluminescence kit (Servicebio, Wuhan, China), and grayscale values were analyzed by ImageJ software. The following antibodies were used: HRP-labeled goat anti-rabbit (1:10,000, ZhongShan JinQiao, ZB-2301, Beijing); HRP-labeled goat anti-mouse (1:10,000, ZhongShan JinQiao, ZB-2305, Beijing); anti-GAPDH (1:50,000, proteintech, 60004-1-IG, United States); anti-HDAC2 (1:1,000, A19626, ABclonal); anti-STAT3 (1:1,000, A1192, ABclonal); anti-p-STAT3 (1:2,000, 9145T, CST); anti-GPX4 (1:1,000, ABclonal, A1933); anti-SLC7A11 (1:1,000, Proteintech, 26864-1-AP, United States); anti-ACSL4 (1:1,000, Abclonal, A6826); anti-CD9 (1:1,000, Abcam, ab92726); anti-TSG101 (1:1,000, Abcam, ab125011).

Following deep anesthesia induced by sodium pentobarbital, the mice were sacrificed, and the hippocampus were obtained and pulverized in a lysis buffer. After centrifugation, the protein concentration of the liquid above was measured using the bicinchoninic acid kit provided by Boster, Ltd., Wuhan, China. Afterward, 20 μg of every nuclear protein sample underwent separation on 10% SDS‐PAGE gels and were then transferred onto a polyvinylidene fluoride membrane. Afterward, the membrane was obstructed using 5% BSA for a duration of 1 h at room temperature. Following this, it was incubated with specific primary antibodies overnight at a temperature of 4°C. Following three washes, the blots were exposed to the suitable secondary antibody for a duration of 1 h at ambient temperature. Immuno‐reactivity was quantified by densitometry using enhanced chemiluminescence.
The antibodies utilized in this study included anti‐GAPDH (A19056; ABclonal) at a dilution of 1:1000, anti‐ACSS2 (A12334; ABclonal) at a dilution of 1:1000, and anti‐HDAC2 (A19626; ABclonal) at a dilution of 1:1000.