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. 2021 Jul 31;13(7):490–512. doi: 10.1007/s13238-021-00864-5

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© The Author(s) 2021

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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LIN28 binds to snoRNA and rRNA, and its loss leads to reduced rRNA biogenesis and ribosomal stress. (A) LIN28A CLIP-seq enrichment analysis. ERVL, MERVL-int and MT2_Mm classes were not enriched in LIN28A binding targets compared with other genes. (B) qRT-PCR showing the expression of rRNA in wild-type and Lin28a knockout ES cells. PCAGIG: empty vector. OE: overexpression. *P < 0.05, ** P < 0.01, *** P < 0.001, two-way ANOVA, n = 3, error bar: standard error of the mean. (C) GSEA analysis showing collective changes in the Ribosome gene set in Lin28a knockout ES cells compared with wild-type cells. NES: normalized enrichment score. (D) Flow cytometry showing OP-puromycin (OP-puro) labeling of wild-type and Lin28a knockout ES cells. (E) Electro-microscopy showing nuclear morphology of wild-type and Lin28a knockout ES cells. Scale bar, 2 μm. (F) Left panel: GSEA analysis showing collective changes in the p53 signaling pathway gene set. Right panel: up-regulated p53 target genes in Lin28a knockout ES cells compared with wild-type cells. (G–I) qRT-PCR showing Dux (G), 2C-related genes (H) or MERVL (I) expression in wild-type and Lin28a knockout ES cells treated with scramble negative control, or Trp53 siRNAs. (J and K) qRT-PCR showing the expression of Dux (J) or 2C-related genes (K) in ES cells treated with 4 μmol/L CX-5461 for 12 h or equal water (control), or in Trp53 knockout ES cells treated with the same conditions. (L) UCSC Genome Browser view of ChIP-seq results using antibodies of p53 and phosphorylated p53 S18P for Dux gene locus. *P < 0.05, **P < 0.01, ***P < 0.001, two-way ANOVA, n = 3, error bar: standard error of the mean