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Vidali et al. (2009) Proc. Natl. Acad. Sci. USA. 106:13341-13346

Rapid formin-mediated actin-filament elongation is essential for polarized plant cell growth.

Movie S1. Functional For2A-3XmEGFP localizes to the apex of growing moss cells. Time-lapse scanning confocal microscopy of a stable line where the For2A locus was modified by integrating 3XmEGFP in frame at the 3 end of the For2A gene. This movie contains 60 images of the medial plane of the cell taken every 20 s, and then compressed into a 6-s QuickTime movie.

Movie S2. Time-lapse acquisition of 1-week-old control RNAi tip cell. Images were acquired with a stereo microscope at 5-min intervals using a 1 lens, zoom 63. To display a representative sequence of a protonemal cell, a subregion was selected, interpolated, and scaled 3 times its original size. (Scale bar, 10 m.)
Movie S3. Time-lapse acquisition of 1-week-old class I RNAi tip cell. Images were acquired with a stereo microscope at 5-min intervals using a 1 lens, zoom 63. To display a representative sequence of a protonemal cell, a subregion was selected, interpolated, and scaled 3 times its original size. (Scale bar, 10 m.)
Movie S4. Profilin/alexa-532-actin polymerization in presence of For2A FH1FH2. Time-lapse TIRF microscopy of moss profilin/actin polymerization in presence of For2A FH1FH2. This movie corresponds to a large field from which the time-lapse series shown in Fig. 6 was taken. Green arrow heads indicate filaments elongating with a rate of 115 subunits/s. Red arrow heads indicate filaments that were elongating rapidly, but then stop. Yellow arrow heads indicate filaments elongating without formin associated at the barbed end, at a rate of 6 subunits/s. Images were taken every 10 s for the indicated time, and then compressed into a 10 frames per second QuickTime movie.

Movie S5. Profilin/Alexa-532-actin polymerization in presence of For1D FH1FH2. Time-lapse TIRF microscopy of moss profilin/actin polymerization in presence of For1D FH1FH2. This movie corresponds to a large field from which the time-lapse series shown in Fig. 6 was taken. Green arrow heads indicate filaments elongating with a rate of 24 subunits/s. Red arrow heads indicate filaments that were elongating with a For1D bound, but then stop. Yellow arrow heads indicate filaments elongating without formin associated at the barbed end, at a rate of 6 subunits/s. Images were taken every 15 s for the indicated time, and then compressed into a 6.6 frames per second QuickTime movie.

Movie S6. Profilin/Alexa-568-actin polymerization in presence of Chim1-2. Time-lapse TIRF microscopy of moss profilin/actin polymerization in presence of Chim1-2. This movie corresponds to the time-lapse series shown in Fig. 6. Green arrow heads indicate filaments elongating with a rate of 43 subunits/s.Yellow arrow heads indicate filaments elongating without formin associated at the barbed end, at a rate of 6 subunits/s. Images were taken every 10 s for the indicated time, and then compressed into a 10 frames per second QuickTime movie.

Movie S7. Profilin/Alexa Fluor 568-actin polymerization in presence of Chim2-1. Time-lapse TIRF microscopy of moss profilin/actin polymerization in presence of Chim2-1. This movie corresponds to a large field from which the time-lapse series shown in Fig. 6 was taken. Images were taken every minute for the indicated time and then compressed into a 1.6 frames per second QuickTime movie.

Movie S8. Profilin/Alexa Fluor 568-actin polymerization without any formin present. Time-lapse TIRF microscopy of moss profilin/actin polymerization. Images were taken every minute for the indicated time, and then compressed into a 1.6 frames per second QuickTime movie.


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