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

. 2023 Feb 20;42:49. doi: 10.1186/s13046-023-02616-1

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

© The Author(s) 2023

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

PMC Copyright notice

Fig. 3 — AQP1 interacted with ANXA2 and recruited it from the cell membrane to the cytoplasm. a Cellular extracts from MDA-MB-231 cells stably transfected Flag-vector or Flag-AQP1 were immunopurified with anti-Flag affinity beads and eluted with Flag peptides. The eluates were resolved on SDS-PAGE and silver-stained followed by mass spectrometry analysis. b-c Immunoprecipitation experiments were performed by using an anti-flag M2 affinity gel and then immunoblotted with ANXA2 or AQP1 antibodies. d Whole cell lysates from Flag-AQP1/MDA-MB-231 cells were immunoprecipitated with anti-ANXA2 or anti-AQP1 and then immunoblotted with antibodies against the indicated proteins. e Immunofluorescence analysis showed the colocalization of AQP1 and ANXA2 in GFP-AQP1/MDA-MB-231 and Flag-AQP1/MDA-MB-231 cells. Scale bar = 20 μm. f Immunohistochemical staining of 194 serial paraffin sections was performed to detect the expression of AQP1 and ANXA2. Scale bar = 100 μm. g Cytoplasmic ANXA2 score in AQP1 high expression group was higher than in AQP1 low expression group in 194 serial paraffin sections (two-tailed Student’s t test, P = 0.0034, ^**P < 0.01). h 70.2% (59/84) IDC patients exhibited high ANXA2 cytoplasmic expression in AQP1 high cytoplasmic expression group, while the percent of high ANXA2 cytoplasmic expression cases was only 33.6% (37/110) in AQP1 low cytoplasmic expression group (Chi-square test, P = 0.000, ^***P < 0.001). i Western blot analysis of ANXA2 expression in Flag-vector/MDA-MB-231 and Flag-AQP1/MDA-MB-231 cells. j Western blot analysis of cytoplasmic ANXA2 expression in the cytosol of Flag-vector/MDA-MB-231 and Flag-AQP1/MDA-MB-231 cells. k Immunofluorescent assay showed the cellular location of ANXA2 in Flag-vector/MDA-MB-231 and Flag-AQP1/MDA-MB-231 cells (n = 194–197 cells per group, Chi-square test, ^***P < 0.001). Scale bar = 20 μm. l Immunohistochemical staining of 194 serial paraffin sections was performed to detect the expression of AQP1 and ANXA2. 21.8% (24/110) IDC patients exhibited ANXA2 membrane expression in AQP1 low cytoplasmic expression group, while the percent of ANXA2 membrane expression cases was only 10.7% (9/84) in AQP1 high cytoplasmic expression group (Chi-square test, P = 0.041, ^*P < 0.05). Scale bar = 100 μm. All in vitro experiments were repeated at least 3 times