a: Immunoblot of α-MYCN immunoprecipitates from MYCN-amplified (IMR32) neuroblastoma cells. Where indicated, EtBr (1 mg/ml) was added to the lysate to disrupt DNA-mediated interactions. The input corresponds to 1 % (n=3).
b: (Left) ChIP of BRCA1 in SH-EP MYCNER cells expressing either shSCR or shUSP11 upon 4-OHT treatment (5 h). Shown is the mean and error bars indicate SD of technical triplicates (n =2). (Right) Immunoblot of cells described above. Actin is loading control (n=2).
c: Immunoblots of indicated proteins after knockdown of USP11 in MYCN-amplified neuroblastoma cells. Asterisk denotes an unspecific band. Tubulin is loading control (n=3).
d: Immunoblots of α-USP11 immunoprecipitates from control cells or cells expressing constitutive MYCN. CDK2 is loading control. The input corresponds to 2 % (n=2).
e: Density plot of MYCN occupancy after 4-OHT treatment (3 h) in control (top) or BRCA1-depleted (bottom) cells with shBRCA1#2 around transcriptional start sites (TSS).
f: Immunoblots of α-USP11 and α-MYCN immunoprecipitates from SH-EP cells stably expressing wildtype (“wt”) MYCN or T58AS62AMYCN (“mut”) as indicated. CDK2 is loading control. The input corresponds to 1 % (n=3).