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

. 2020 Oct 1;11:4931. doi: 10.1038/s41467-020-18708-x

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

© The Author(s) 2020

Open Access This 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 3 — a MAGE-A11-induced ubiquitination of PCF11 depends on its substrate recognition by MAGE-A11 SBC. Ubiquitinated proteins from FLAG-vector, FLAG-MAGE-A11 wild-type, or SBC mutants stably expressing HEK293FT cells were treated with 10 µM MG132 for 4 h and isolated with tandem ubiquitin binding entity-agarose followed by SDS-PAGE and immunoblotting for endogenous PCF11. Source data are provided as a Source Data file. b MAGE-A11 SBC mutants fail to promote proliferation of MAGE-A11 knockout DAOY cells. Wild-type, MAGE-A11-knockout DAOY cells, or those reconstituted with MAGE-A11 were counted for cell proliferation at the indicated time points (n = 3 biologically independent experiments per group). Data are mean ± SEM. p-value by two-way ANOVA followed by Tukey’s multiple comparisons test is indicated (****P < 0.0001). c–e MAGE-A11 knockout DAOY cells reconstituted with MAGE-A11 SBC mutants reduced clonogenic growth and xenograft tumor growth of MAGE-A11 knockout DAOY cells. Stable expression of wild-type MAGE-A11 or SBC mutants in MAGE-A11-knockout DAOY cells were assayed for clonogenic growth (c) (n = 3 biologically independent experiments per group, two-tailed unpaired Student’s t test, *P < 0.05; **P < 0.01), Data are mean ± SEM and xenograft tumor growth in mice (n = 6 per group). Tumor burden (d) and tumor weight (e) are shown. Data are mean ± SD. p-value by two-way ANOVA followed by Tukey’s multiple comparisons test is indicated in d (****P < 0.0001), ordinary one-way ANOVA followed by Sidak’s multiple comparisons test is indicated in e (**P < 0.01, ***P < 0.001, ****P < 0.0001). Red circle, Vector; Blue square, MAGE-A11 WT; Green triangle, MAGE-A11 F275A; and Pink triangle, MAGE-A11 M341R. f MAGE-A11 M341R mutant fails to promote mRNA 3’UTR shortening. Transcriptome analysis of tumors (n = 3 per group) from MAGE-A11 knockout DAOY cells stably expressing FLAG-MAGE-A11 wild-type or M341R mutant was determined and scatterplot of Percentage of Distal polyA site Usage Index (PDUI) is shown. False discovery rate ≤ 0.05 and ΔPDUIs ≥ 0.2 are colored. The shift towards distal polyadenylation site is significant (p = 5.9 × 10⁻¹⁶, binomial test). g Representative RNA-seq density plots are shown.