Western blot analysis of neuritin expression
For Western blot analysis, proteins in the cell extracts were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) through an 8 % polyacrylamide gel and were then transferred onto a nitrocellulose membrane. The membrane was incubated with 5 % non-fat milk in PBS and later with anti-neuritin MAb for 2 h at room temperature. After washing, the membranes were incubated with an alkaline phosphatase-conjugated goat anti-mouse IgG antibody (Amersham Biosciences, Buckinghamshire, England) for 1 h at room temperature. Immunoreactive bands were detected using the ECL Western blot analysis system (Amersham Biosciences, Buckinghamshire, England).