JCS02860 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Fluorescence micrographs of dilution-induced actin filament disassembly reactions. Unlabeled actin filaments were diluted at time zero (from 5 μM to 0.1 μM) in the presence of different factors at pH 5.0: control buffer (Actin alone), 5 μM Lat-A, 100 μM Twf1, 1 μM CP, or 100 μM Twf1 with 1 μM CP. At the indicated time points, samples were removed from reactions, incubated with Alexa Fluor 488-phalloidin and visualized by light microscopy. Bar, 5 μm.

• Movie 1 -

  Movie 1. TIRF microscopy of fluorescently labeled actin filaments incubated with control buffer. This movie corresponds to the time-lapse series shown in Fig. 4A.

• Movie 2 -

  Movie 2. TIRF microscopy of fluorescently labeled actin filaments incubated with 10 μM Lat A. This movie corresponds to the time-lapse series shown in Fig. 4B.

• Movie 3 -

  Movie 3. TIRF microscopy of fluorescently labeled actin filaments incubated with 2 μM Twf1. This movie corresponds to the time-lapse series shown in Figs 4C and 4D. Upon addition of fresh actin monomers to the flow cell (following severing of filaments by Twf1), yellow arrowheads track the growth of barbed ends, whereas blue arrowheads are kept stationary at the original positions of barbed ends.

• Movie 4 -

  Movie 4. TIRF microscopy of fluorescently labeled actin filaments incubated with 2 μM Twf1 and 3 μM CP. This movie corresponds to the time-lapse series shown in Figs 6D and 6E. Upon addition of fresh actin monomers to the flow cell (following rare severing of filaments by Twf1 in the presence of CP), yellow arrowheads track the growth of barbed ends, whereas blue arrowheads are kept stationary at the original positions of barbed ends.

• Movie 5 -

  Movie 5. TIRF microscopy of fluorescently labeled actin filaments incubated with 2 μM Twf1-10. This movie corresponds to the time-lapse series shown in Figs 7C and 7D. Upon addition of fresh actin monomers to the flow cell (following severing of filaments by Twf1-10), yellow arrowheads track the growth of barbed ends, whereas blue arrowheads are kept stationary at the original positions of barbed ends.

• Movie 6 -

  Movie 6. TIRF microscopy of fluorescently labeled actin filaments incubated with 2 μM Twf1-10 and 3 μM CP. This movie corresponds to the time-lapse series shown in Figs 7E and 7F. Upon addition of fresh actin monomers to the flow cell (following severing of filaments by Twf1-10 in the presence of CP), yellow arrowheads track the growth of barbed ends, whereas blue arrowheads are kept stationary at the original positions of barbed ends.