Western blot tests were performed as previously reported [30 (link)]. Briefly, whole left hemispheres of rats in different groups were isolated and collected 24 h after SAH. After protein extraction, equal amounts of protein samples were loaded onto each lane of SDS-PAGE gel. After electrophoresis, the protein samples were transferred onto a nitrocellulose membrane for blocking and incubation at 4 °C overnight with the following primary antibodies: Anti-osteopontin (1:1000, sc-21742), anti-ERK 1/2 (phosphor-Thr202/Tyr204) (1:200, sc-16982), anti-ERK1/2 (1:2000, sc- 514302), and anti-β-actin (1:5000, I-19) from Santa Cruz Biotechnology Inc., TX, USA; anti-Beclin 1 (1:1000, NB500-249), anti-ATG 5 (1:500, NB110-53818), anti-LC3 (1:5000, NB600-1384) from Novus Biologicals, CO, USA; anti-FAK (phospho-Y397) (1:1000, ab81298), anti-FAK (1:1000, ab131435), anti-SQSTM1/p62 (1:5000, ab56416) from Abcam, Cambridge, MA, USA). Secondary antibodies used were anti-mouse (1:5000, sc-2031), anti-goat (1:5000, sc-2354) from Santa Cruz Biotechnology Inc., and anti-rabbit (1:5000, 2839792) from EMD Millipore Corporation, MA, USA.
Free full text: Click here
