C myc

The extraction of total RNA and reverse transcription into cDNA were the same as described above. The specific primers for the fusion gene were 5'-CCTACCCTCTCAACGACAGC-3' (forward) and 5'-CTCTGACCTTTTGCAAGGAG-3' (reverse). Those for β-actin were the same as the above. PCR condition was as follow: pre-denaturation at 95°C for 5 min; 35 cycles of denaturation at 95°C for 30 s, annealing at 56°C for 30 s and extension at 72°C for 45s; a final extension at 72°C for 5 min. The products were electrophoresed on 1.5% agarose gel and analyzed with Quantity One Software (BIO-RAD, USA).
Protein concentrations in the cell lysates were determined by BCA protein assay kit (Bi Yun Tian, Shang Hai). Equal amounts of proteins (50 μg) were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and blotted onto nitrocellulose sheets. The proteins were then probed with the mouse monoclonal antibody C-MYC (Zhongshan Goldenbridge Biotechnology Co., Ltd., Beijing, China) and goat anti-mouse secondary antibodies (Zhongshan Goldenbridge Biotechnology Co., Ltd., Beijing, China). Protein bands were visualized using the Quantity One Software (BIO-RAD, USA).