Protein expressions in the ipsilateral hemisphere were measured by western blot as previously described [3 (link)]. Protein samples with equal volumes were loaded on an SDS-PAGE gel, and then electrophoresed protein bands were transferred to a nitrocellulose membrane. The membrane was incubated with the following primary antibodies overnight at 4 °C: rabbit polyclonal anti-frizzled 7 (1:1000; ab64636, Abcam, MA), rabbit polyclonal anti-WISP1 (1:2000; ab178547, Abcam, MA), mouse monoclonal anti-Dvl (1:200; sc-166303, Santa Cruz Biotechnology, TX), rabbit monoclonal anti-β-Catenin (1:10,000; ab32572, Abcam, MA), rabbit polyclonal anti-phospho-β-Catenin (Ser33/37/Thr41) (1:1000; #9561, Cell Signaling, MA), rat monoclonal anti-ZO-1 (1:200, sc-33725, Santa Cruz Biotechnology, TX), rabbit polyclonal anti-Claudin-5 (1:500; ab15106, Abcam, MA), and mouse monoclonal anti-VE-Cadherin (1:500; sc-9989, Santa Cruz Biotechnology, TX). Appropriate secondary antibody (Santa Cruz Biotechnology, TX) was applied to the membranes and incubated for 1 h at room temperature. Immunoblots were probed and exposed to films. Band densities were analyzed using Image J software (NIH, Bethesda, MD).
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