Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

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

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 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/.

PMC Copyright notice

LIN28A is associated with nucleolar proteins and mediates rRNA biogenesis and 2C gene repression through the NCL/TRIM28 complex. (A) Venn diagrams showing the overlap between proteins pulled down by LIN28A and nucleolar proteins or ribosomal proteins. (B) Validation of the interaction between the endogenous LIN28A and nucleolar or nuclear proteins by Co-IP followed by Western blot analysis in ES cells. IgG was used as a negative control for the IP. (C) RIP-qPCR assays for NCL and FBL in wild-type and Lin28a knockout ES cells. *P < 0.05, **P < 0.01, ***P < 0.001, two-way ANOVA, n = 3, error bar: standard error of the mean. (D) Quantification of 2’-O-Me by RiboMethseq for all bases in 28s, 18s and 5s rRNAs in wild-type and Lin28a knockout ES cells. MethScore was calculated using the method mentioned in (Birkedal et al., 2015; Marchand et al., 2016). ***P < 0.001. Wilcox signed rank test. (E) Left panel: RiboMeth-seq showing profiles of mapped sequencing reads in a range of a snoRNA targeting region on the 28s rRNA (from 3,366 bp to 3,397 bp). Red highlighted A is a predicted methylation site (from SnoRNA Orthological Gene Database). Right panel, calculated RiboMeth-Seq score to indicate the level of the 2’-O-methylation at the predicted site of A (Birkedal et al., 2015). (F) Proteomics analysis showing the nucleus and cytoplasm proportion of ribosomal proteins in wild-type and Lin28a knockout ES cells. ***P < 0.001, t-test, n = 3, error bar: standard error of the mean. (G) Proteomics analysis showing the relative abundances of indicated proteins in wild-type and Lin28 knockout cells (averaged proteomics data from Lin28a knockout ES cells and Lin28a/b knockout iPS cells). Error bar: standard error of the mean. n = 2 independent proteomics experiments. (H and I) qRT-PCR showing the expression of 2C-related genes in ES cells treated with scramble negative control or the indicated siRNAs. (J and K) NCL and TRIM28 ChIP-qPCR analysis at the Dux (J) and rRNA (K) loci in wild-type and Lin28a knockout ES cells. (L) p53 and TRIM28 ChIP-qPCR analysis at the Dux loci in untreated and CX-5461-treated ES cells