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

. 2019 Oct 7;9:14401. doi: 10.1038/s41598-019-50955-x

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

© The Author(s) 2019

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

Transwell migration experiments of ABMSC, FBMSC and HUVEC in response to serum. (A) Representative widefield immunofluorescence (IF) images of ABMSC, FBMSC and HUVEC at the top (left panel) and bottom (right panel) of a Transwell filter. Cells were allowed to migrate for 24 hours and were subsequently fixed and stained for Lamin A/C (green), F-actin (red). Pore structures (8 µm) were visualized by DIC imaging. Scalebar: 40 μm. (B) Schematic overview of Transwell experiments. During transmigration, different migratory stages can be distinguished. Stage 1: cells on top of the Transwell filter that have not started transmigration. Stage 2: cells on top of the Transwell filter that form protrusions through the filter pores and initiate transmigration. Stage 3: cells that have completed transmigration through the pores. (C) Quantification of the percentage of transmigrated ABMSC, FBMSC and HUVEC after 4 and 24 hours migration. Percentage is determined from IF tilescan widefield images stained for Dapi and Lamin A/C from both top and bottom comprising the total transwell area. Median ± range. n = 3 independent experiments using cells from 3 different donors. **p < 0.01 (Kruskal-Wallis, multiple comparisons uncorrected Dunn’s test). (D) Quantification of total F-actin at the bottom of the Transwell. F-actin surface area coverage at top and bottom of the Transwell filters was determined from IF tilescan widefield images comprising the total transwell area. Cell-body migration (F-actin at Transwell bottom (stage2 + 3) was calculated as a percentage of total F-Actin (all stages combined). Median ± range. n = 3 independent experiments using cells from 3 different donors. ns = non-significant, *p < 0.05 (Kruskal-Wallis, multiple comparisons uncorrected Dunn’s test). (E) Quantification of the percentage of protruding cells that are fully transmigrated. Widefield images from the bottom of the filter were manually screened for presence of a nucleus (Lamin A/C) and presence of cell-body (F-Actin). Number of fully transmigrated cells (stage 3) was calculated as percentage of total protruding cells (stage 2 + 3). Median ± range. n ≥ 3 independent experiments including 30 cells each. *p < 0.05, **p < 0.01 (Kruskal-Wallis, multiple comparisons uncorrected Dunn’s test).