*2.7. Immunoblot Analysis*

Cells were harvested with a lysis bu ffer, and the extracted proteins were separated by SDS polyacrylamide gel electrophoresis (7.5–10%) and electrophoretically transferred to a polyvinylidene fluoride membrane. The transblots were preincubated with 5% nonfat dry skim milk or BSA in Tris-bu ffered saline (TBS, pH 7.4) and then incubated overnight with the antibody against each protein. After washing with TBS/0.05% Tween 20, the membranes were incubated with either anti-rabbit, anti-mouse, or anti-goat IgG HRP-linked antibody (1:3000). The membrane was rinsed with TBS/0.05% Tween 20, and the immunoreactive bands were developed by ECL systems (Millipore Billerica, MA, USA).
