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 Oct 17;13:6128. doi: 10.1038/s41467-022-33683-1

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. 1 — a, Top: Image of a cell stained for NHE1 showing preferential localization at the leading edge (yellow arrowhead). Bottom: Image of another cell showing intense localization of SWELL1-GFP at the cell rear (white arrowhead). b Front to rear ratio of (i) endogenous NHE1 (n = 48) or (ii) SWELL1-GFP intensity (n = 23) in confined cells. Data represent mean ± SD from four independent experiments. c Western blots of cells transduced with SC or shRNA sequences against NHE1 and/or SWELL1. GAPDH served as a loading control. Uncropped blots in Source Data. d Effects of NHE1 and/or SWELL1 knockdown on cell volume inside confining channels. Data represent mean ± SD for cells analyzed from three independent experiments. e, f Effects of NHE1 and/or SWELL1 knockdown on e migration velocity and f cell entry time in confining channels. Data represent mean ± SD for cells analyzed from 3 independent experiments. g Images showing dissemination of SC and dual NHE1- and SWELL1-KD cells from spheroids embedded in 3D collagen gels at t = 0 and 5 h. h–k Effects of NHE1 and/or SWELL1 knockdown on h the time for the first cell to dissociate from spheroids, and the i migration velocity, j mean squared displacement, and k trajectories of disseminated cells in 3D. Data represent mean ± SD for cells analyzed from three independent experiments. l, m Effects of NHE1 and/or SWELL1 knockdown on l normalized area of expansion at t = 12 h relative to t = 0 and m circularity of the spheroids at t = 0 embedded in 3D collagen gels. Data represent mean ± SD for cells analyzed from three independent experiments. n Time-lapse montage of a SWELL1-GFP-tagged cell (outlined by dashed magenta lines) dissociating from a spheroid embedded in collagen. White arrowheads denote SWELL1 polarization at the cell rear. Yellow arrowheads indicate the cell leading edge. **p < 0.01 and ***p < 0.001 relative to SC, ^###p < 0.001 relative to either of single KD cells. Tests performed: d, e, h, i, m one-way ANOVA followed by Tukey’s post hoc test, f, l Kruskal–Wallis followed by Dunn’s, or j two-way ANOVA followed by Tukey’s. The number of cells analyzed is indicated in each panel. Cell model: MDA-MB-231.