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. 2009 Dec 30;285(9):6835–6847. doi: 10.1074/jbc.M109.068155

FIGURE 4.

FIGURE 4.

Severing activity of TgADF as observed by TIRF microscopy. A, severing of actin filaments by TgADF and S. pombe cofilin. Fluorescence time-lapse micrographs of actin filaments were taken over a period of 0–12 min after the addition of 0.3 μm S. pombe cofilin (SpCOF, top), 0.3 μm TgADF (middle), or 1.5 μm TgADF (bottom) at time zero. Rabbit actin co-polymerized with Alexa Fluor 488-labeled actin was tethered to flow chambers with N-ethylmaleimide-treated myosin. Time-lapse TIRF microscopy was used to visualize filament severing by TgADF and S. pombe cofilin over time. Scale bar, 10 μm. B, quantitation of the rate of filament disassembly by TgADF and S. pombe cofilin. The average length (mean ± S.E.) of the 15 longest filaments in the field of view was calculated at the indicated time points after TgADF or SpCOF addition and plotted for each condition (n = 3 experiments). C, detailed montage of actin filaments being severed by 1.5 μm TgADF over time. Scale bar, 10 μm.