Fig. 2

IGF2BP1 stimulated MYCN expression is 3´UTR-, miRNA- and BTYNB-dependent. a Western blot (n = 6) and RT-qPCR (n = 5) analysis of MYCN expression upon IGF2BP1-KO (I1-KO) in BE(2)-C. b MYCN mRNA decay monitored by RT-qPCR in control (grey) and I1-KO (blue) BE(2)-C upon indicated time of actinomycin D (ActD) treatment (n = 3). c IGF2BP1-RIP analyses in parental BE(2)-C (n = 5). d Western blotting of miTRAP from BE(2)-C lysates using MS2-fused MYCN 3´UTR or MS2-RNA (n = 1). e BTYNB-response curve in BE(2)-C (n = 3). f IGF2BP1-RIP in BE(2)-C treated with DMSO or BTYNB (n = 3). g Western blot analysis of MYCN expression in BE(2)-C treated with BTYNB (BTY, n = 3) for 3 and 6 d or replacement of BTY by DMSO after 3 d treatment. Treatment scheme in lower panel. h AGO2-RIP analyses in I1-KO versus control BE(2)-C (n = 3). i Scheme of luciferase reporters constructs. j Expression of miRNAs in BE(2)-C (left) and mutated fraction of MYCN-targeting miRNA-binding sites (right). k, l Activity of indicated luciferase reporters in control and I1-KO (k, n = 5) or DMSO- and BTYNB-treated BE(2)-C (l, n = 3)