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

. 2021 Aug 19;32(18):1772–1791. doi: 10.1091/mbc.E21-04-0174

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

© 2021 Adams et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License.

PMC Copyright notice

FIGURE 5: — Analysis of the effects of the overexpression of actin regulatory proteins on leader bleb formation and cell migration. A375M melanoma cells cultured on BSA (1 µg/ml)-treated coverslips with confinement by a PDMS pad resting on 3 µm beads to define the confinement height and induce leader bleb–based motility. Cells were transiently coexpressing various mEmerald (mEm)-tagged actin regulatory proteins or GFP control together with FusionRed-F-Tractin. Analysis (A–I) presented for cells in which the nucleus was localized in the bleb; see Supplemental Figure 5 for analysis of cells with the nucleus in the cell body. (A–I) Fusion proteins expressed are color coded: GFP control (white); actin assembly-regulating proteins (red, mEmerald-ARP3, mEmerald-cortactin, mEmerald-mDia1, mEmerald-VASP); actomyosin contraction-regulating proteins (green, mEmerald-non-muscle myosin IIA, mEmerald non-muscle-myosin IIB, mEmerald-tropomyosin 2, mEmerald-tropomyosin 4); actin cross-linking proteins (blue, mEmerald-α2-spectrin, mEmerald-α-actinin-1, mEmerald-fascin-1, mEmerald-filamin-A. Average length (A) and width (B) of leader bleb and cell motility speed (E); n (cells) is shown below categories. Error is SEM. Statistical significance was determined by one-way ANOVA. **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001, NS not significant. (C) Rose plots of representative migration tracks (8 h, 10 min intervals) of cells overexpressing the noted fusion protein and undergoing LBBM; the number of cells is noted on each plot. (D) Fraction of cells undergoing leader bleb–based motility with the nucleus localized in the cell body vs. the bleb. The number of cells analyzed (n) is shown below each category. Average mean squared displacement over time for cells undergoing LBBM and overexpressing (F) aII spectrin (G) a-actinin (H) fascin-1 or (I) filamin-A; n (cells) = 29 (a-2-spectrin), 42 (α-actinin-1, 34 (fascin-1), and 37 (filamin-A) cells.