Rabbit anti plexin a1

The following antibodies were used: mouse Tau-1 (Chemicon, MAB3420, 1:200), mouse anti-MAP2 (SigmaAldrich, M4403, 1:1500), rabbit anti-MAP2 (Abcam, ab32454, 1:1000), mouse anti-Ankyrin-G (Antibodies Inc., 75–146, 1:100), rabbit anti-Plexin-A1 (Abcam, ab23391, 1:1000), mouse anti-GFP (Covance, MMS-118P, 1:1000), mouse anti-FLAG M2 (SigmaAldrich, F3165, 1:1000), anti-Rap1 (Upstate, #07–916, 1:200), anti-Sema3A (Abcam, ab23393, 1:200), SMI-312 (BioLegend, 837904, 1:200) and goat secondary antibodies labeled with Alexa-350 (Molecular Probes, 1:200), −488 (1:800) or −594 (1:800). Nuclei were stained with Hoechst 33342 (Molecular Probes, 1:6000).

The cold RIPA buffer (Thermo Scientific, Rockford, IL, USA) was utilized for cell lysis. Minute (TM) Cytoplasmic and Nuclear Fractionation kit (SC003) was used to extract nucleoprotein. Thereafter, we used the BCA Protein Assay kit (Beijing Solarbio Science & Technology Co., Ltd., china) for quantifying protein content. After separating proteins through SDS-PAGE, they were transferred on the PVDF membranes. Then, each membrane was subjected to overnight incubation using primary monoclonal antibodies, including mouse anti-VEGFR2, rabbit anti-plexinA1 and mouse anti-gapdh (all Abcam, Cambridge, MA, UK), rabbit anti-Histon H3, rabbit anti-JAK, rabbit anti-STAT3, rabbit anti-p-JAK and rabbit anti-p-STAT3 (all ABclonal Co., Ltd,Wuhan, China) as well as mouse anti-β-actin (Cell Signaling Technology, Danvers, MA, USA) antibodies under 4°C. Afterwards, HRP-labeled secondary antibodies (Santa Cruz, CA, USA) were further used to incubate the membranes for 2 h. In addition, the enhanced chemiluminescence (ECL) substrate (Pierce, Rockford, IL, USA) was used for detecting protein blots, whereas the Image analysis software was employed for visualization.