Figure 3
From: Upstream ORF affects MYCN translation depending on exon 1b alternative splicing
Translation of uORF and existence of MYCNOT. a) SH-EP cells were transfected with either p-uORF/MYCN (lane 1) or p-uORF/MYCN Δ1b(lane 2). Western blot was conducted on 40 μg proteins, 48 hours after transfection with -upper panel: a monoclonal antibody against the V5 epitope (1:300) and -lower panel: the anti β-Actin antibody (1:10000). L: protein ladder; b) SH-EP cells were transfected with p-uORF/MYCN Δ1b. Western blot was conducted on 40 μg protein, 48 hours after transfection with -left panel: a monoclonal antibody against the V5 epitope (1:300) and -right panel: a polyclonal rabbit anti-NGAERSPQSPAGRRA peptide [anti-MYCNOT] polyclonal antibody (1:100) L: protein ladder; c) Western blot analysis of 40 μg protein extracts from adult (lane 1) and foetal (lane 2) brain. Results obtained with -upper panel: the anti-MYCNOT antibody (1:100) and -lower panel: the anti β-Actin antibody (1:10000).