Anti sox11

Equal amount of total proteins from each sample were separated with SDS-PAGE gel and transferred onto nitrocellulose membrane with the Trans-blot SD semi-dry transfer cell (Bio-Rad, Hercules, CA). The membrane was blocked with 5% non-fat dry milk (Santa Cruz Biotech, Dallas, TX) for one hour, and then incubated with anti-SOX11 (Santa Cruz Biotech, #Sc-20,096), anti-SDCCAG8 (GeneTex, Irvine, CA, # GTX115484) or anti-GAPDH (GeneTex, #GTX100118) primary antibody, followed by incubation with HRP-conjugated secondary antibody (GE Healthcare, Piscataway, NJ). Signal detection was performed with the ECL-Plus Kit (GE Healthcare) and protein bands were quantified with the NIH Image J.

Total protein was lysed from cells using radioimmunoprecipitation assay buffer (RIPA buffer; Beyotime, Shanghai, China) supplemented with protease inhibitors. SDS/PAGE was used to separate 30 µg of total proteins, which were then transferred to a nitrocellulose membrane. The membranes were blocked with 5% non-fat milk and then incubated overnight at 4°C with specific primary antibodies. The membranes were washed and incubated with secondary antibodies at room temperature for 1 h. Electro chemiluminescent detection was used to visualize the protein bands. The expressions of GAPDH and β-actin were used as internal controls. The primary antibodies were used in this study: anti-SOX11 (Santa Cruz, CA, United States), anti-MYCN (#84406, Cell Signaling Technology, United States), anti-p21 (ab109199, Abcam, United States), anti-p27 (ab32034, Abcam, United States), anti-Cyclin D1 (ab134175, Abcam, United States), anti-NSE (ab180943, Abcam, United States), anti-CHGA (ab68271, Abcam, United States), and anti-SYP (ab32127, Abcam, United States). Quantification of the western blots was performed using ImageJ.