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. 2019 Feb 26;151(5):660–669. doi: 10.1085/jgp.201812199

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© 2019 Solís and Russell

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/).

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Figure 3. — Stiffness and PKC signaling increase CapZ FRAP dynamics that is diminished by PIP2 reduction, while β-tentacle-truncated CapZ is irresponsive to stimuli. (A and B) FRAP kinetics were measured on NRVMs plated on PAA substrates (10 kPa and 100 kPa) or glass and infected with CapZβ1-GFP (A) or CapZβ1ΔC-GFP virus (B). FRAP kinetics in untreated (UT; white bars) NRVMs or NRVMs treated with 500 µM neomycin for 30 min to reduce the PIP2 level (Neo; gray bars) or PMA for 30 min (black bars). (A) k_FRAP is higher on 100-kPa substrates and glass relative to 10-kPa substrates (insert). k_FRAP was low on 10-kPa substrates and not reduced further by neomycin. However, k_FRAP on stiffer substrates (100 kPa) or glass were initially higher but significantly reduced by neomycin. PMA increased k_FRAP on stiff substrates. (B) CapZβ1ΔC-GFP dynamics is insensitive to stiffness, neomycin, and PMA treatments (mean ± SEM, n = 6–16 in A and n = 4–9 in B; *, P < 0.05, ***, P < 0.01).