Western Blot Analysis of Protein Targets

Cells or brain tissues were collected and lysed with lysis buffer (Beyotime) containing protease inhibitor (Roche). The protein concentration was measured using the Bradford assay (Bio-Rad). The lysates samples with equal amounts of total protein were separated by SDS-PAGE electrophoresis (PAGE) and then transferred to polyvinylidene fluoride membranes (PVDF; Millipore). After blocking with 5% skim milk-TBST for 1 h, the indicated targets were probed with primary antibodies and appropriate peroxidase-conjugated secondary antibodies. The antibodies used in the experiments include anti-IE1 [61 (link)] and anti-pUL97 (gift from William J. Britt, University of Alabama, USA)[62 (link)], anti-IE1/2 (p1215, Virusys), anti-pp65 (p1205, Virusys), anti-UL44 (p1202-1, Virusys), anti-pp28 (CA004-1,Virusys), anti-mIE1 [63 (link)], anti-GAPDH (10494-1-AP, Protientech), anti-FLAG (20543-1-AP, Protientech), anti-RFX7 (NBP1-71819, Novus), anti-SOCS3 (ab16030, Abcam), anti-GFAP (16825-1-AP, Proteintech), anti-Nestin (19483-1-AP, Proteintech), anti-DCX (13925-1-AP, Proteintech), anti-SOX2 (ab97959, Abcam), Phospho-(Ser/Thr) Substrate Antibody (9611S, Cell Signaling Technology), anti-STAT3 (10253-2-AP, Proteintech) and anti-Phospho-STAT3 (Tyr705) (9145, Cell Signaling Technology). The protein bands were detected using a Chemiluminescence machine and analyzed by a densitometry program (Image J).