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( A ) Increasing CapZ concentrations shift condensates from toroidal morphologies (0 to 50 nM) to rod-like morphologies at intermediate concentrations (125 to 250 nM) and back to spheres at high CapZ concentrations (1000 nM). Scale bars, 5 μm. ( B to E ) Quantification of the (B) fraction of condensates with aspect ratios greater than 1.2, (C) average aspect ratio, (D) fraction of condensates displaying toroidal morphologies, and (E) histogram of all condensate aspect ratios for conditions in (A). Condensates with aspect ratios > 20 (0, 0.3, 0.6, 0.5, and 0% of data for the 0, 50, 125, 250, and 1000 nM conditions) were hidden to better visualize the distributions. ( F ) Increasing <t>gelsolin</t> concentrations shift condensates from toroid-dominated morphologies to rod-like condensate morphologies. Scale bars, 5 μm. ( G to J ) Quantification of the (G) fraction of condensates that display a toroidal morphology, (H) fraction of condensates that have aspect ratios greater than 1.2, (I) average condensate aspect ratio, and (J) histogram of all condensate aspect ratio distributions for the conditions in (F). Condensates with aspect ratios > 20 (corresponding to 1.3, 1.9, 2.6, and 0.59% of data for the 0, 10, 50, and 100 nM conditions) were hidden to better visualize the distributions. ( K ) Cartoon depicting the hypothesized mechanism by which CapZ and gelsolin inhibit trapped ring and toroid formation and restore rod-like condensate morphologies. For all bar charts, data are the means across three independent experiments with more than 500 condensates analyzed per condition. Overlaid gray circles denote the means of each replicate. * P < 0.05, ** P < 0.01, and *** P < 0.001 using unpaired, two-tailed t tests on replicate means ( n = 3). Conditions were 10 μM VASP condensates with 3 μM Mg 2+ -actin and 3 <t>μM</t> <t>profilin</t> to provide toroid-dominated conditions with the indicated concentration of capping factor.
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( A ) Increasing CapZ concentrations shift condensates from toroidal morphologies (0 to 50 nM) to rod-like morphologies at intermediate concentrations (125 to 250 nM) and back to spheres at high CapZ concentrations (1000 nM). Scale bars, 5 μm. ( B to E ) Quantification of the (B) fraction of condensates with aspect ratios greater than 1.2, (C) average aspect ratio, (D) fraction of condensates displaying toroidal morphologies, and (E) histogram of all condensate aspect ratios for conditions in (A). Condensates with aspect ratios > 20 (0, 0.3, 0.6, 0.5, and 0% of data for the 0, 50, 125, 250, and 1000 nM conditions) were hidden to better visualize the distributions. ( F ) Increasing <t>gelsolin</t> concentrations shift condensates from toroid-dominated morphologies to rod-like condensate morphologies. Scale bars, 5 μm. ( G to J ) Quantification of the (G) fraction of condensates that display a toroidal morphology, (H) fraction of condensates that have aspect ratios greater than 1.2, (I) average condensate aspect ratio, and (J) histogram of all condensate aspect ratio distributions for the conditions in (F). Condensates with aspect ratios > 20 (corresponding to 1.3, 1.9, 2.6, and 0.59% of data for the 0, 10, 50, and 100 nM conditions) were hidden to better visualize the distributions. ( K ) Cartoon depicting the hypothesized mechanism by which CapZ and gelsolin inhibit trapped ring and toroid formation and restore rod-like condensate morphologies. For all bar charts, data are the means across three independent experiments with more than 500 condensates analyzed per condition. Overlaid gray circles denote the means of each replicate. * P < 0.05, ** P < 0.01, and *** P < 0.001 using unpaired, two-tailed t tests on replicate means ( n = 3). Conditions were 10 μM VASP condensates with 3 μM Mg 2+ -actin and 3 <t>μM</t> <t>profilin</t> to provide toroid-dominated conditions with the indicated concentration of capping factor.
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( A ) Increasing CapZ concentrations shift condensates from toroidal morphologies (0 to 50 nM) to rod-like morphologies at intermediate concentrations (125 to 250 nM) and back to spheres at high CapZ concentrations (1000 nM). Scale bars, 5 μm. ( B to E ) Quantification of the (B) fraction of condensates with aspect ratios greater than 1.2, (C) average aspect ratio, (D) fraction of condensates displaying toroidal morphologies, and (E) histogram of all condensate aspect ratios for conditions in (A). Condensates with aspect ratios > 20 (0, 0.3, 0.6, 0.5, and 0% of data for the 0, 50, 125, 250, and 1000 nM conditions) were hidden to better visualize the distributions. ( F ) Increasing <t>gelsolin</t> concentrations shift condensates from toroid-dominated morphologies to rod-like condensate morphologies. Scale bars, 5 μm. ( G to J ) Quantification of the (G) fraction of condensates that display a toroidal morphology, (H) fraction of condensates that have aspect ratios greater than 1.2, (I) average condensate aspect ratio, and (J) histogram of all condensate aspect ratio distributions for the conditions in (F). Condensates with aspect ratios > 20 (corresponding to 1.3, 1.9, 2.6, and 0.59% of data for the 0, 10, 50, and 100 nM conditions) were hidden to better visualize the distributions. ( K ) Cartoon depicting the hypothesized mechanism by which CapZ and gelsolin inhibit trapped ring and toroid formation and restore rod-like condensate morphologies. For all bar charts, data are the means across three independent experiments with more than 500 condensates analyzed per condition. Overlaid gray circles denote the means of each replicate. * P < 0.05, ** P < 0.01, and *** P < 0.001 using unpaired, two-tailed t tests on replicate means ( n = 3). Conditions were 10 μM VASP condensates with 3 μM Mg 2+ -actin and 3 <t>μM</t> <t>profilin</t> to provide toroid-dominated conditions with the indicated concentration of capping factor.
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( A ) Increasing CapZ concentrations shift condensates from toroidal morphologies (0 to 50 nM) to rod-like morphologies at intermediate concentrations (125 to 250 nM) and back to spheres at high CapZ concentrations (1000 nM). Scale bars, 5 μm. ( B to E ) Quantification of the (B) fraction of condensates with aspect ratios greater than 1.2, (C) average aspect ratio, (D) fraction of condensates displaying toroidal morphologies, and (E) histogram of all condensate aspect ratios for conditions in (A). Condensates with aspect ratios > 20 (0, 0.3, 0.6, 0.5, and 0% of data for the 0, 50, 125, 250, and 1000 nM conditions) were hidden to better visualize the distributions. ( F ) Increasing <t>gelsolin</t> concentrations shift condensates from toroid-dominated morphologies to rod-like condensate morphologies. Scale bars, 5 μm. ( G to J ) Quantification of the (G) fraction of condensates that display a toroidal morphology, (H) fraction of condensates that have aspect ratios greater than 1.2, (I) average condensate aspect ratio, and (J) histogram of all condensate aspect ratio distributions for the conditions in (F). Condensates with aspect ratios > 20 (corresponding to 1.3, 1.9, 2.6, and 0.59% of data for the 0, 10, 50, and 100 nM conditions) were hidden to better visualize the distributions. ( K ) Cartoon depicting the hypothesized mechanism by which CapZ and gelsolin inhibit trapped ring and toroid formation and restore rod-like condensate morphologies. For all bar charts, data are the means across three independent experiments with more than 500 condensates analyzed per condition. Overlaid gray circles denote the means of each replicate. * P < 0.05, ** P < 0.01, and *** P < 0.001 using unpaired, two-tailed t tests on replicate means ( n = 3). Conditions were 10 μM VASP condensates with 3 μM Mg 2+ -actin and 3 <t>μM</t> <t>profilin</t> to provide toroid-dominated conditions with the indicated concentration of capping factor.
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( A ) Increasing CapZ concentrations shift condensates from toroidal morphologies (0 to 50 nM) to rod-like morphologies at intermediate concentrations (125 to 250 nM) and back to spheres at high CapZ concentrations (1000 nM). Scale bars, 5 μm. ( B to E ) Quantification of the (B) fraction of condensates with aspect ratios greater than 1.2, (C) average aspect ratio, (D) fraction of condensates displaying toroidal morphologies, and (E) histogram of all condensate aspect ratios for conditions in (A). Condensates with aspect ratios > 20 (0, 0.3, 0.6, 0.5, and 0% of data for the 0, 50, 125, 250, and 1000 nM conditions) were hidden to better visualize the distributions. ( F ) Increasing <t>gelsolin</t> concentrations shift condensates from toroid-dominated morphologies to rod-like condensate morphologies. Scale bars, 5 μm. ( G to J ) Quantification of the (G) fraction of condensates that display a toroidal morphology, (H) fraction of condensates that have aspect ratios greater than 1.2, (I) average condensate aspect ratio, and (J) histogram of all condensate aspect ratio distributions for the conditions in (F). Condensates with aspect ratios > 20 (corresponding to 1.3, 1.9, 2.6, and 0.59% of data for the 0, 10, 50, and 100 nM conditions) were hidden to better visualize the distributions. ( K ) Cartoon depicting the hypothesized mechanism by which CapZ and gelsolin inhibit trapped ring and toroid formation and restore rod-like condensate morphologies. For all bar charts, data are the means across three independent experiments with more than 500 condensates analyzed per condition. Overlaid gray circles denote the means of each replicate. * P < 0.05, ** P < 0.01, and *** P < 0.001 using unpaired, two-tailed t tests on replicate means ( n = 3). Conditions were 10 μM VASP condensates with 3 μM Mg 2+ -actin and 3 <t>μM</t> <t>profilin</t> to provide toroid-dominated conditions with the indicated concentration of capping factor.
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( A ) Increasing CapZ concentrations shift condensates from toroidal morphologies (0 to 50 nM) to rod-like morphologies at intermediate concentrations (125 to 250 nM) and back to spheres at high CapZ concentrations (1000 nM). Scale bars, 5 μm. ( B to E ) Quantification of the (B) fraction of condensates with aspect ratios greater than 1.2, (C) average aspect ratio, (D) fraction of condensates displaying toroidal morphologies, and (E) histogram of all condensate aspect ratios for conditions in (A). Condensates with aspect ratios > 20 (0, 0.3, 0.6, 0.5, and 0% of data for the 0, 50, 125, 250, and 1000 nM conditions) were hidden to better visualize the distributions. ( F ) Increasing <t>gelsolin</t> concentrations shift condensates from toroid-dominated morphologies to rod-like condensate morphologies. Scale bars, 5 μm. ( G to J ) Quantification of the (G) fraction of condensates that display a toroidal morphology, (H) fraction of condensates that have aspect ratios greater than 1.2, (I) average condensate aspect ratio, and (J) histogram of all condensate aspect ratio distributions for the conditions in (F). Condensates with aspect ratios > 20 (corresponding to 1.3, 1.9, 2.6, and 0.59% of data for the 0, 10, 50, and 100 nM conditions) were hidden to better visualize the distributions. ( K ) Cartoon depicting the hypothesized mechanism by which CapZ and gelsolin inhibit trapped ring and toroid formation and restore rod-like condensate morphologies. For all bar charts, data are the means across three independent experiments with more than 500 condensates analyzed per condition. Overlaid gray circles denote the means of each replicate. * P < 0.05, ** P < 0.01, and *** P < 0.001 using unpaired, two-tailed t tests on replicate means ( n = 3). Conditions were 10 μM VASP condensates with 3 μM Mg 2+ -actin and 3 <t>μM</t> <t>profilin</t> to provide toroid-dominated conditions with the indicated concentration of capping factor.
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( A ) Increasing CapZ concentrations shift condensates from toroidal morphologies (0 to 50 nM) to rod-like morphologies at intermediate concentrations (125 to 250 nM) and back to spheres at high CapZ concentrations (1000 nM). Scale bars, 5 μm. ( B to E ) Quantification of the (B) fraction of condensates with aspect ratios greater than 1.2, (C) average aspect ratio, (D) fraction of condensates displaying toroidal morphologies, and (E) histogram of all condensate aspect ratios for conditions in (A). Condensates with aspect ratios > 20 (0, 0.3, 0.6, 0.5, and 0% of data for the 0, 50, 125, 250, and 1000 nM conditions) were hidden to better visualize the distributions. ( F ) Increasing <t>gelsolin</t> concentrations shift condensates from toroid-dominated morphologies to rod-like condensate morphologies. Scale bars, 5 μm. ( G to J ) Quantification of the (G) fraction of condensates that display a toroidal morphology, (H) fraction of condensates that have aspect ratios greater than 1.2, (I) average condensate aspect ratio, and (J) histogram of all condensate aspect ratio distributions for the conditions in (F). Condensates with aspect ratios > 20 (corresponding to 1.3, 1.9, 2.6, and 0.59% of data for the 0, 10, 50, and 100 nM conditions) were hidden to better visualize the distributions. ( K ) Cartoon depicting the hypothesized mechanism by which CapZ and gelsolin inhibit trapped ring and toroid formation and restore rod-like condensate morphologies. For all bar charts, data are the means across three independent experiments with more than 500 condensates analyzed per condition. Overlaid gray circles denote the means of each replicate. * P < 0.05, ** P < 0.01, and *** P < 0.001 using unpaired, two-tailed t tests on replicate means ( n = 3). Conditions were 10 μM VASP condensates with 3 μM Mg 2+ -actin and 3 <t>μM</t> <t>profilin</t> to provide toroid-dominated conditions with the indicated concentration of capping factor.
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( A ) Increasing CapZ concentrations shift condensates from toroidal morphologies (0 to 50 nM) to rod-like morphologies at intermediate concentrations (125 to 250 nM) and back to spheres at high CapZ concentrations (1000 nM). Scale bars, 5 μm. ( B to E ) Quantification of the (B) fraction of condensates with aspect ratios greater than 1.2, (C) average aspect ratio, (D) fraction of condensates displaying toroidal morphologies, and (E) histogram of all condensate aspect ratios for conditions in (A). Condensates with aspect ratios > 20 (0, 0.3, 0.6, 0.5, and 0% of data for the 0, 50, 125, 250, and 1000 nM conditions) were hidden to better visualize the distributions. ( F ) Increasing <t>gelsolin</t> concentrations shift condensates from toroid-dominated morphologies to rod-like condensate morphologies. Scale bars, 5 μm. ( G to J ) Quantification of the (G) fraction of condensates that display a toroidal morphology, (H) fraction of condensates that have aspect ratios greater than 1.2, (I) average condensate aspect ratio, and (J) histogram of all condensate aspect ratio distributions for the conditions in (F). Condensates with aspect ratios > 20 (corresponding to 1.3, 1.9, 2.6, and 0.59% of data for the 0, 10, 50, and 100 nM conditions) were hidden to better visualize the distributions. ( K ) Cartoon depicting the hypothesized mechanism by which CapZ and gelsolin inhibit trapped ring and toroid formation and restore rod-like condensate morphologies. For all bar charts, data are the means across three independent experiments with more than 500 condensates analyzed per condition. Overlaid gray circles denote the means of each replicate. * P < 0.05, ** P < 0.01, and *** P < 0.001 using unpaired, two-tailed t tests on replicate means ( n = 3). Conditions were 10 μM VASP condensates with 3 μM Mg 2+ -actin and 3 <t>μM</t> <t>profilin</t> to provide toroid-dominated conditions with the indicated concentration of capping factor.
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( A ) Increasing CapZ concentrations shift condensates from toroidal morphologies (0 to 50 nM) to rod-like morphologies at intermediate concentrations (125 to 250 nM) and back to spheres at high CapZ concentrations (1000 nM). Scale bars, 5 μm. ( B to E ) Quantification of the (B) fraction of condensates with aspect ratios greater than 1.2, (C) average aspect ratio, (D) fraction of condensates displaying toroidal morphologies, and (E) histogram of all condensate aspect ratios for conditions in (A). Condensates with aspect ratios > 20 (0, 0.3, 0.6, 0.5, and 0% of data for the 0, 50, 125, 250, and 1000 nM conditions) were hidden to better visualize the distributions. ( F ) Increasing gelsolin concentrations shift condensates from toroid-dominated morphologies to rod-like condensate morphologies. Scale bars, 5 μm. ( G to J ) Quantification of the (G) fraction of condensates that display a toroidal morphology, (H) fraction of condensates that have aspect ratios greater than 1.2, (I) average condensate aspect ratio, and (J) histogram of all condensate aspect ratio distributions for the conditions in (F). Condensates with aspect ratios > 20 (corresponding to 1.3, 1.9, 2.6, and 0.59% of data for the 0, 10, 50, and 100 nM conditions) were hidden to better visualize the distributions. ( K ) Cartoon depicting the hypothesized mechanism by which CapZ and gelsolin inhibit trapped ring and toroid formation and restore rod-like condensate morphologies. For all bar charts, data are the means across three independent experiments with more than 500 condensates analyzed per condition. Overlaid gray circles denote the means of each replicate. * P < 0.05, ** P < 0.01, and *** P < 0.001 using unpaired, two-tailed t tests on replicate means ( n = 3). Conditions were 10 μM VASP condensates with 3 μM Mg 2+ -actin and 3 μM profilin to provide toroid-dominated conditions with the indicated concentration of capping factor.

Journal: Science Advances

Article Title: A balance between nucleating and elongating actin filaments controls deformation of protein condensates

doi: 10.1126/sciadv.aea0196

Figure Lengend Snippet: ( A ) Increasing CapZ concentrations shift condensates from toroidal morphologies (0 to 50 nM) to rod-like morphologies at intermediate concentrations (125 to 250 nM) and back to spheres at high CapZ concentrations (1000 nM). Scale bars, 5 μm. ( B to E ) Quantification of the (B) fraction of condensates with aspect ratios greater than 1.2, (C) average aspect ratio, (D) fraction of condensates displaying toroidal morphologies, and (E) histogram of all condensate aspect ratios for conditions in (A). Condensates with aspect ratios > 20 (0, 0.3, 0.6, 0.5, and 0% of data for the 0, 50, 125, 250, and 1000 nM conditions) were hidden to better visualize the distributions. ( F ) Increasing gelsolin concentrations shift condensates from toroid-dominated morphologies to rod-like condensate morphologies. Scale bars, 5 μm. ( G to J ) Quantification of the (G) fraction of condensates that display a toroidal morphology, (H) fraction of condensates that have aspect ratios greater than 1.2, (I) average condensate aspect ratio, and (J) histogram of all condensate aspect ratio distributions for the conditions in (F). Condensates with aspect ratios > 20 (corresponding to 1.3, 1.9, 2.6, and 0.59% of data for the 0, 10, 50, and 100 nM conditions) were hidden to better visualize the distributions. ( K ) Cartoon depicting the hypothesized mechanism by which CapZ and gelsolin inhibit trapped ring and toroid formation and restore rod-like condensate morphologies. For all bar charts, data are the means across three independent experiments with more than 500 condensates analyzed per condition. Overlaid gray circles denote the means of each replicate. * P < 0.05, ** P < 0.01, and *** P < 0.001 using unpaired, two-tailed t tests on replicate means ( n = 3). Conditions were 10 μM VASP condensates with 3 μM Mg 2+ -actin and 3 μM profilin to provide toroid-dominated conditions with the indicated concentration of capping factor.

Article Snippet: Rabbit muscle actin (catalog no. AKL95), human gelsolin (catalog no. HPG6), and human profilin 1 (catalog no. PR02) were purchased from Cytoskeleton.

Techniques: Two Tailed Test, Concentration Assay