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 Jan 8;219(2):e201904137. doi: 10.1083/jcb.201904137

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

© 2020 Wang et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

Figure 1. — Inverse correlation between integrin β4 expression and FA maturation. (A) Immunofluorescence confocal images of migrating PA-JEB/β4 keratinocytes stained for BP230 (green), integrin β4 (Itg. β4; red), phosphorylated paxillin (PAX-pY31; blue), phosphorylated MLC (MLC-pS19; green), vinculin (Vinc; red), and actin (Act; blue) 72 h after creating the gap. Nuclei were counterstained with DAPI (cyan). T means the trailing area, while L means the leading area of migrating cell monolayer. Yellow arrowheads indicate the enrichment of hemidesmosomal structure at the leading border, and red arrowheads indicate the higher level of MLC-pS19 and larger FAs in the leader cells. Scale bars: 20 µm. For quantification, type I HD (based on β4 and BP230 colocalization), MLC-pS19, and vinculin-positive areas are calculated as a percentage of the total ROI area. The values represent the mean (± SD) of three independent experiments, with ∼18 images per experiment. ***, P < 0. 001; ****, P < 0.0001. (B) Representative confocal fluorescence microscopy images of PA-JEB (β4 −) and PA-JEB/β4 (β4 +) keratinocytes cultured for 1 d in complete KGM medium and then switched to DMEM (10% FCS) for 16 h. Cells were immunostained for β4 (green), plectin (Plec; red), and laminin-332 (Ln332; blue). Colocalization of β4, plectin, and laminin-332 is visualized in the overlay images. Nuclei were counterstained with DAPI (cyan). (C) Cells were immunostained for β4 (green), vinculin (Vinc; red), and actin (Act; blue). Nuclei were counterstained with DAPI (cyan). Scale bars: 10 µm. (D) Cell area and FA size probed by vinculin were quantified with ImageJ. The values represent the mean (± SD) of three independent experiments, with ∼20 images per experiment. ****, P < 0.0001.