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. 2021 Jul 15;220(9):e202005214. doi: 10.1083/jcb.202005214

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© 2021 Cowell et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 4. — TLNRD1 is an actin-bundling protein.(A) Schematic of talin ABS2 binding a single actin filament (left). TLNRD1-FL (middle) and TLNRD1-4H (right) bind two actin filaments. (B and C) Actin bundling assays using serial dilutions of TLNRD1-FL (B) and TLNRD1-4H (C) against 15 µM F-actin. (D–F) EM images of negative-stained F-actin bundles with (D) TLNRD1-FL, (E) TLNRD1-4H, and (F) TLNRD1-F250D. Red box (left) enlarged on right. (G) U2OS cells expressing GFP-TLNRD1 were plated on fibronectin, incubated for 2 h with SiR-actin to label the actin cytoskeleton, and imaged live using an Airyscan confocal microscope (1 picture every 16 s; Video 1). Scale bars: (main) 25 µm, (inset) 5 µm. The yellow arrows highlight GFP-TLNRD1 localizing to actin stress fibers.