(A) Venn diagrams of LIN28B-bound (gray) and transcripts with significant differential polysome enrichment (blue and red) as well as LIN28B-bound and transcriptional targets of MYCN (yellow). (B) Gene ontology analysis (GO Biological Process 2018) of indicated overlaps (LIN28B-bound and polysome-enriched transcripts [left] and LIN28B-bound and MYCN targets [right]). (C) Scatterplot depicting transcripts depending on their polysome enrichment and LIN28B binding. MYCN targets of the ribosome are indicated in red. (D) Protein synthesis analysis measured by metabolic labeling using S35-labeled methionine in the respective groups (n = 6 biological replicates per group). (E) In vitro translation assay with RPS29-reporter mRNA on lysates of BE2C-KO cells expressing the different LIN28B proteins: WT and deficient in RNA binding (MUT), only intact zinc knuckle domain (ZKD), and only intact cold shock domain (CSD) (n = 3 biological replicates). Differential comparisons were made between BE2C CTRL and BE2C LIN28B-KO cells. MYCN targets were defined using CHIPseeker (promoter ≤1kb). Statistical data in D were assessed using 2-tailed Student’s t test, in E using 1-way ANOVA with Tukey’s multiple-comparison test; data are presented as mean ± SEM. *P < 0.05; ****P < 0.0001. GO analysis in B was performed with Enrichr. MYCN CHIP-Seq data were derived from Durbin et al. (74).