*3.4. Western Blot Analysis*

Cells were lysed on ice for 30 min in 100 ΐL lysis buffer (120 mM NaCl, 40 mM Tris (pH 8), and 0.1% NP-40) and centrifuged at 13,000× *g* for 15 min. Supernatants were collected and the protein concentrations were determined. Aliquots containing 40 ΐg of protein were boiled for 5 min and electrophoresed on 10% SDSpolyacrylamide gels. Proteins were transferred to nitrocellulose membranes, which were subsequently incubated with a primary antibody overnight at 4 °C. The membranes were further incubated with horseradish peroxidase-conjugated antiimmunoglobulin-G (Pierce, Rockford, IL, USA). Protein bands were detected using an enhanced chemiluminescence western blotting detection kit (Amersham Biosciences, Little Chalfont, Buckinghamshire, UK). 
