Fig. 6: Effects of cofilin on various actin densities.

a Maximum intensity projection of actin networks after 10 min illumination (upper panels) and 10 min after stopping the illumination (lower panels). Dense condition: 5 μM Actin, 15 μM Profilin, 50 nM Arp2/3 complex, 25 nM CP, 150 nM SspB-mScarlet-I-VCA, and 300 nM Cofilin. Sparse condition: 2 μM Actin, 6 μM Profilin, 50 nM Arp2/3 complex, 10 nM CP, 150 nM SspB-mScarlet-I-VCA, and 300 nM Cofilin. All scale bars, 20 μm. b Box plots of the fluorescence intensities of Alexa647-actin after 10 min illumination (left boxes) and 10 min after stopping the illumination (right boxes) in the presence of 300 nM cofilin. n = 6 and 8 for dense and sparse conditions, respectively. p values were calculated by unpaired two-sided t-test. **p < 0.01. c Relationship between the initial fluorescence intensity of Alexa647-actin and the actin disassembly level. The disassembly level was calculated by dividing the intensity 10 min after illumination by the intensity 10 min after stopping the illumination. Bold lines indicate power-law fitting. n = 51, 60, 57, 42 for 0, 100, 300, 900 nM Cofilin, respectively. d Estimation of the actin network density. Each color corresponds to the range of actin density in indicated illumination powers. e Spatial distribution of cofilin in the dense condition. Condition: 5 μM Actin, 15 μM Profilin, 100 nM Arp2/3 complex, 25 nM CP, 150 nM SspB-mScarlet-I-VCA, and 900 nM Alexa546-cofilin. Scale bar, 20 μm. f Relationship between the fluorescence intensities of actin and cofilin in single actin networks. n = 28 from individual actin networks. g Three-dimensional reconstruction of floating actin networks on 6% DGS-NTA(Ni) lipid at each indicated time point produced by various illumination patterns. h Three-dimensional reconstruction of floating actin networks by a square-shaped illumination pattern on 6% DGS-NTA(Ni) lipid at each indicated time point. Yellow and white arrowheads indicate the first and second polymerization, respectively. g, h Condition: 5 μM Actin, 15 μM Profilin, 100 nM Arp2/3 complex, 25 nM CP, 150 nM SspB-mScarlet-I-VCA, and 600 nM Cofilin. Source data are provided as a Source Data file.