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 Nov 4;9:15911. doi: 10.1038/s41598-019-52040-9

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

Rho/ROCK signal is essential for fibre formation induced by Prom1. (A) The inhibitors targeting the small GTPases (100 nM of EHT1864 for Rac, 50 µM of ZCL278 for Cdc42, 20 µM of Y-27632 for Rho and Rac, 0.5 µg/ml of C3 for Rho, 10 μM of Blebbistatin for Myosin II) were treated for 6 hours and the expression plasmid conveying Prom1-YFP was transfected. Cells were analysed with GFP antibody or phalloidin at 24 hpt. (B,C) Quantitative data for (A). The numbers (B) and lengths (C) of the fibres were counted and measured, respectively. The experiments were repeated four times, in each of which 20 cells were analysed. Data represent mean ± SE of these four experiments. (D) The activation of RhoA is essential for fibre formation by Prom1. The plasmid conveying myc-tagged dominant-negative version of RhoA (dn-RhoA) was co-transfected with Prom1-FL. Staining was performed with GFP (for Prom1), myc (for dn-RhoA) antibodies and phalloidin. Outlined arrowheads represent the cell that got the transfection of Prom1, but not dn-Rho. (E) The active Rho coincides with Prom1 in the membrane extensions. The plasmid conveying myc-tagged constitutively-active version of RhoA (ca-RhoA) was co-transfected with Prom1-FL. Staining was performed with GFP (for Prom1), myc (for dn-RhoA) antibodies and phalloidin. Enlarged images corresponding to the white squares are shown in the bottom three panels. (F) The fibres are formed at the point where Prom1 and active-Rho encounter with each other. The plasmids conveying GFP-rGBD and Prom1-mCherry (F) or control-mCherry (G) were co-transfected and time-lapse imaging was performed for 6 minutes at 24 hpt, focusing on the initial points of the fibre formation. Yellow and white arrowheads indicate the points with high (where Rho is active) and low GFP (where Rho is inactive) intensities, respectively. 16 fields were analysed, and representative images are shown. See also Supplementary Movie S2. Scale bars, 10 µm (A,E), 1 µm (two right panels), 20 µm (D), 1 µm (F,G).