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 Jun 17;10:9868. doi: 10.1038/s41598-020-66789-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

LXN forms a complex with HECTD1 and Rps3 in HIEC cell. (A,B) Cell lysates from HIEC cells were immunoprecipitated with anti-LXN antibody or control IgG, and then the purified protein complex was separated by 10% SDS-PAGE and stained with coomassie brilliant blue (A), the differential bands were cut and in-gel digested with trypsin. Peptides were subject to LC-MALDI MS/MS assay. Identified peptide and MS/MS spectrum that matched with the sequence of HECTD1 and Rps3 are shown (B). (C) HEK293T cells were co-transfected with HA-HECTD1 and Flag-LXN plasmids. 48 h after co-transfection, immunoprecipitation was performed with either anti-Flag antibody or anti-HA antibody to detect the interaction of HECTD1 with LXN. (D) Cell lysates obtained from HIEC cells were immunoprecipitated with anti-LXN, anti-HECTD1 antibody, respectively, IgG as control antibody, and then separated by 10% SDS-PAGE. Transferred membrane was immunoblotted with antibodies as indicated. (E,F) HCT116 cells were cultured in 6-well plates. The localization of HECTD1 and LXN (E), Rps3 and LXN (F) were analyzed by immunofluorescence staining with HECTD1, Rps3 and LXN antibody. Higher magnification inserts show details of localization. The pixel intensity of localization in the selected regions (white line) are shown. Scale bars = 10 μm. (G) HA-HECTD1 expression vector in combination with either empty vector or expression vectors of Flag-LXN mutants were co-transfected into HEK293T cells. Extracted proteins were precipitated by anti-HA antibody, and then separated by 10% SDS-PAGE. The transferred membrane was immunoblotted with either anti-Flag or anti-HA antibody.