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

. 2024 Jan 2;43(2):250–276. doi: 10.1038/s44318-023-00018-9

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

© The Author(s) 2024

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 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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

PMC Copyright notice

Figure EV3 — (A–D) HEK293T cells were transfected with the indicated constructs: (A) HA-mutant PHOX2B (+13Ala). (B) HA-mutant RUNX2 (+12Ala). (C) HA-mutant HOXD13 (+10Ala). (D) HA-mutant PABPN1 ( + 7 Ala). Endogenous UBA6 was immunoprecipitated from the nuclear fraction (Nuc, LaminB1 enriched) or the cytoplasmic (Cyt, GAPDH enriched) fraction (unrelated IgG was used as a control). The immunocomplexes were analyzed with anti-HA antibodies. (E) HEK293T cells were transfected with constructs expressing different polyalanine-expanded disease proteins (mutant PHOX2B, mutant RUNX2, mutant HOXD13, and mutant PABPN1) together with FLAG-USE1. Cell lysates were incubated without β mercaptoethanol and analyzed for ubiquitin loading. Results are mean ± s.e.m. normalized to control (empty vector, no disease protein). Paired 2-tailed t test. n = 3–6 biological replicates. (F) Control and UBA6-depleted HEK293T cells were transfected with HA-Ub, mutant PHOX2B or empty vector and incubated for the last 6 h with the proteasome inhibitor MG132 (10 μM). Endogenous E6AP was immunoprecipitated from cell lysates for ubiquitination analysis. (G) HEK293T cells were transfected with WT PHOX2B, mutant PHOX2B or empty vector and incubated for the last 6 h with the proteasome inhibitor MG132 (10 μM). Endogenous E6AP was immunoprecipitated from cell lysates for ubiquitination analysis (unrelated IgG was used as a control). ns non-significant, *P < 0.05, **P < 0.01.