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. 2019 Jun 10;17(6):e3000317. doi: 10.1371/journal.pbio.3000317

Fig 2. Modulation of the branched-to-linear actin network balance by the Arp2/3 complex.

Fig 2

The underlying data can be found within S1 Data. A. Fluorescence snapshots of actin networks assembled around WASp-coated microbeads in the presence of fluorescent actin, profilin, capping protein, and variable concentrations of Arp2/3 complex. Images were taken 30 min after the initiation of the experiment. Scale bar: 5 μm. B. Fluorescence snapshots of actin networks assembled around formin-coated microbeads in the presence of fluorescent actin, profilin, capping protein, and variable concentrations of Arp2/3 complex. Images were taken 30 min after the initiation of the experiment. Scale bar: 5 μm. C. Quantification of (A). Rate of actin assembly around WASp-coated microbeads as a function of the Arp2/3 complex concentration, normalized to the maximum value. D. Quantification of (B). Rate of actin assembly around formin-coated microbeads as a function of the Arp2/3 complex concentration, normalized to the maximum value. E. Snapshots of the actin cytoskeleton organization in budding yeast cells fixed and labeled with fluorescent phalloidin, in the presence or in the absence of 200 μM CK-666 (left images), and for the formin defective mutant bni1-FH2#1 bnr1Δ at nonrestrictive temperature (25 °C; center images) or restrictive temperature (37 °C; right images). Scale bars: 2 μm. F. Quantification of (E). Average number of actin patches and cables per cell. G. In vitro deviation index, calculated as a function of the Arp2/3 complex concentration. This index compares how actin assembly rates around WASp and formin-coated beads deviate from a balanced situation in which both types of networks assemble optimally. H. In vivo deviation index, based on structures number, calculated in the presence of DMSO, 200 μM CK-666, at 37 °C for wild-type cells or at 37 °C for bni1-FH2#1 bnr1Δ cells. This index compares how the number of actin patches and cables deviate from the wild-type condition. Arp2/3, actin-related protein 2/3; CK-666, Arp2/3 complex inhibitor I; WASp, Wiskott–Aldrich syndrome protein.