2.14. Western blot analysis for PC12 cells

The PC12 cells were stimulated for equivalent periods of time and lysed in extraction buffer (10 mM Tris [pH 7.4], 150 mM NaCl, 1% Triton X-100, 5 mM EDTA [pH 8.0]). The protein samples were separated by SDS-PAGE (10%) and electro-transferred onto a nitrocellulose (NC) membrane (Boster Biotechnology Co., Ltd., China). Protein expression was evaluated using antibodies against P53, Bax, Bcl-2, NGFR and α-syn (Boster Biotechnology Co., Ltd., China). The blots were incubated with the appropriate secondary antibodies conjugated to alkaline phosphatase (AP) peroxidase (Boster Biological Technology Co., Ltd., China). The protein levels were normalized by re-probing the blots with antibody against β-actin (Boster Biological Technology Co., Ltd., China) as shown in Figure ​Figure22e,f.