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

. 2015 Aug 24;212(9):1465–1479. doi: 10.1084/jem.20150384

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

© 2015 Inoue et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

PMC Copyright notice

Figure 4. — CNOT3-mediated posttranscriptional regulation of p53 mRNA. (A) Relative expression levels of spliced p53 mature mRNA and unspliced pre-mRNA in bKO pro–B cells compared with control pro–B cells, as determined by real-time qPCR. Error bars represent SD. n = 4 biological replicates. RT(−), no reverse transcription. (B) Stability of p53 mature mRNA. Pro–B cells from control and bKO mice were treated with actinomycin D for the indicated periods of time, and the relative mRNA levels compared with time 0 were determined by real-time qPCR. Data are shown as mean ± SD of n = 3 biological replicates. (C) Association of CNOT1 and CNOT3 with the 3′-UTR of p53 mRNA. FLAG-tagged 3′-UTRs of p53 and Gapdh mRNA were incubated with lysates of an Abelson virus–transformed pro–B cell line and immunoprecipitated with anti-FLAG antibody, followed by Western blotting. (D) Association of CNOT3 with p53 mRNA by RIP assay. Wild-type pro–B cell lysates were immunoprecipitated with anti-CNOT3 antibody or control IgG, and the immunoprecipitates were analyzed by RT-PCR with primers specific to p53 and Gapdh mRNA. (E) Poly(A) tail length of p53 mRNA in pre-pro–B and pro–B cells of control and bKO mice was determined by the LM-PAT assay. (F) Flow cytometry of intracellular p53 protein expression in pro–B cells from control, bKO, and p53^−/− mice. Numbers indicate the mean fluorescence intensity of each population. Data are representative of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; Student’s t test.