G Technology) at 1:1,000) in 2.5 milk. The next day, the membrane was rinsed with Tris-buffered saline (137 mM NaCl, 2.7 mM KCl, 19 mM Tric Base, pH 7.4) containing 0.1 Tween-20 and incubated with secondary anti-rabbit and anti-mouse antibodies (1:20,000 for both) conjugated with horseradish peroxidase. Immunoblots were visualized by chemiluminescence (GE Healthcare) and recorded on Fujifilm LAS