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

. 2022 Feb 5;41(12):1752–1766. doi: 10.1038/s41388-022-02203-2

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

© The Author(s) 2022

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. 2 — A Co-immunoprecipitation (IP) and immunoblotting (IB) analysis of the Erk3-EGLN3 interaction. Total cell lysates prepared from HEK293T cells transfected with the indicated plasmids were immuoprecipitated with anti-Flag or control mouse IgG and then immunoblotted with anti-myc. B GST pulldown analysis of the interaction between Erk3 and EGLN3 or its fragments. Total cell lysates were incubated with Glutathione-Sepharose 4B beads and then immunoblotted with anti-Flag. C Schematic representation of full-length or different fragments of EGLN3. D GST pulldown analysis of the interaction between EGLN3 and Erk3 or its fragments. E Schematic representation of full-length or different fragments of Erk3. F Cycloheximide chase experiment was conducted to analyze the effect of EGLN3 or EGLN3R205K on Erk3 stability. HEK293T cells were transfected with the indicated plasmids and then treated with CHX (30 µg/mL) for the indicated times. Total cell lysates were analyzed by IB with the indicated antibodies. G–J HEK293T cells transfected with the indicated plasmids (G–I) or bone marrow-derived macrophages (J) were treated with Chloroquine (200 µM, 8, 12, and 16 h, respectively), Bafilomycin A1 (100 ng/mL, 16 h), or MG132 (10 µM, 8 h). IB was conducted with the indicated antibodies. GST glutathione S-transferase; IgH heavy chain of IgG; mIg mouse immunoglobulin; IP immunoprecipitation; IB immunoblotting; FL full-length; CHX cycloheximide; BMDM bone marrow-derived macrophages; PD pull-down; Ab antibody; Baf Bafilomycin A1; WT wild-type; KI knock-in.Shown are representative images of three independent experiments.