Ab227560

Western blot was performed as previously reported.^{19 (link)} Briefly, the rats were sacrificed at 3h, 6h, 12h, 24h, and 72h after SAH. Rats were anesthetized and perfused with 0.1M PBS, and the left hemispheres were immediately harvested. Samples were mixed with RIPA, after centrifuged at 14000g for 20min (4°C), the supernatants were collected. A protein assay kit was used for measuring the protein concentration of the samples. Equal amounts of proteins (40μg) were loaded onto the SDS-PAGE gel and transferred onto polyvinylidene fluoride membranes. Membranes were incubated with primary antibodies against EZH2 (1;1000, ab191080, Abcam, USA), H3K27Me3 (1:1000,ab6002, Abcam, USA), H3 (1:1000, ab1791, Abcam, USA), SOCS3(1:1000, ab14939, Abcam, USA), TRAF6 (1:1000, ab227560, Abcam, USA), NF-κB(1:1000, ab16502, Abcam, USA), IL-1β (1:1000, ab9722, Abcam, USA), IL-6 (1:1000, ab6672, Abcam, USA), TNF-α (1:1000, ab1793, Abcam, USA), and IL-10 (1:1000, ab33471, Abcam, USA) overnight at 4°C. On the following day, secondary antibodies were incubated at room temperature for 2h. The bands were visualized by ECL reagents. The proteins band densities were analyzed with Image J software.