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Volume 7 issue 12 December 2006
Dynamic organization of the actin cytoskeleton during meiosis and spore formation in budding yeast
Christof Taxis, Celine Maeder, Simone Reber, Nicole Rathfelder, Kota Miura, Klaus Greger, Ernst H.K. Stelzer and Michael Knop
Supplemental Figure S1: Actin organization in mitotic and meiotic cells in wild type and Abp140p-4GFP cells. Actin was visualized using rhodamine-phalloidin, DNA using Hoechst 33342 staining. The size of the bar is 4 µm. (i) wild-type cell during mitotic cell division. (ii) ABP140::4GFP cell during mitotic cell division. (iii) wild-type cell during meiosis I. (iv) ABP140::4GFP cell during meiosis I. (v) wild-type cell during meiosis II. (vi) ABP140::4GFP cell during meiosis II.
Figure S1 (.jpg)
Supplemental Movie S1: The movie shows a Don1p-GFP expressing cell undergoing PSM assembly. Maximum projections from image stacks taken every minute are used for the assembly of the movie. The movie corresponds to the frames from Figure 1B. Movie plays at 1 frame per 0.1 sec.
Movie S1 (.mov)
Supplemental Movie S2: The movie shows a Don1p-GFP expressing cell undergoing PSM growth and closure. Movie recording conditions as for Movie S1. The movie corresponds to the frames from Figure 1B. Movie plays at 1 frame per 0.1 sec.
Movie S2 (.mov)
Supplemental Movie S3: The movie shows a Don1p-GFP expressing cell in meiosis during the time point of PSM assembly. Maximum projections from image stacks taken every 4 sec. are used for the assembly of the movie. The first frame of the movie shows two arrows pointing to the two precursor structures, for which merging was observed (Figure 1C). Movie plays at 1 frame per 0.1 sec.
Movie S3 (.mov)
Supplemental Movie S4: As for Movie S3, but for the cell shown in Figure 1D.
Movie S4 (.mov)
Supplemental Movie S5: Precursor movement in Δmpc54 Δmpc70 and Don1p-GFP expressing cells in meiosis. Recording conditions as for Movie S3. To the right of the movie is a processed version using a walking average of 4 frames. This movie was generated using ImageJ and the walking average plugin. This allows better visualization of fast movements. The movie corresponds to Figure 3A.
Movie S5 (.mov)
Supplemental Movie S6: The movie shows a montage of four cells from two different stages of PSM assembly. In the cells in an early phase (two cells on the top), the Don1p-GFP staining is associated with precursors in the cytoplasm and at the SPBs (indicated). The four SPBs can easily be distinguished from other precursors, because the move in a pair wise manner, which is due to the two short meiosis II spindle that connect them in pairs. In later stages (cells at the bottom), Don1p-GFP localizes to the LEP coat (indicated). Control cells (two cells to the left) and Lat-A treated cells are shown. The movie corresponds to Figure 3C and D.
Movie S6 (.mov)
Supplemental Movie S7: Time-lapse movie of Abp140p-4GFP and Don1p-RFP expressing cells undergoing meiosis. The movie shows 12 time points during meiosis, each reperesented by a full turn of the cell. The data stacks were recorded every 15 minutes. Each turn was created from projections (10° intervals) using SoftWorxTM of a data stack recorded at the respective time point. Don1-RFP (red colour) labelled PSM precursors, LEP coats (donut stage) and diffuse Don1-RFP staining (after closure of the PSMs) and actin cables (Abp140-4GFP, green colour) can be identified. The movie corresponds to Figure 5A.
Movie S7 (.mov)
Supplemental Movie S8: Actin cable organization in meiosis. The movie shows a 360° sourround view of two Abp140-4GFP expressing cells during meiosis. The data stack was recorded using a SPIM (selective plane illumination microscope (57) ) and deconvolved. Projections were created every 5° using ImageJ. The movie corresponds to Figure 5B.
Movie S8 (.mov)
Supplemental Movie S9 and S10: Visualization of actin cable dynamics in meiosis using Abp140p-4GFP cells. The movie shows a single plane from the periphery of a cell. Pictures were taken every 96 msec. The movie corresponds to Figure 5C and D.
Movie S9 (.mov) and Movie S10 (.mov)
Supplemental Movie S11: Movement of a short actin cable. Movie shows maximum projections from three planes recorded from the periphery of a cell spanning a section of approx 1,5 µm. Frames were taken every 820 msec. The movie corresponds to Figure 5E.
Movie S11 (.mov)
Supplemental Movie S12: Movement of a short actin cable in a cell treated with a low dose of Lat-A. Movie recording as for Movie S11. The movie corresponds to Figure 5E.
Movie S12 (.mov)
Supplemental Movie S13: Movement of a Don1p-RFP (red colour) labelled precursor along an actin cable in a Δmpc54 cell in meiosis II. The actin cable was visualized using Abp140p-4GFP (green colour). The frames were taken every 2 seconds. The movie corresponds to Figure 5F.
Movie S13 (.mov)
Supplemental Movie S14: Time lapse movie of GFP-Cap2p and Don1p-RFP expressing cells undergoing meiosis. The movie shows 12 time points during meiosis, each reperesented by a full turn of the cell. The data stacks were recorded every 15 minutes. Each turn was created from projections (10° intervals) of a data stack recorded at the respective time point. Don1-RFP (red colour) labelled PSM precursors, LEP coats (donut stage) and actin patches (GFP-Cap2p, green colour) can be identified. The transition from precursor to donut stage is shown. The movie corresponds to Figure 6A (i) and (ii).
Movie S14 (.mov)
Supplemental Movie S15: Same as Movie S14, but the transition from donut stage to closed PSM is shown.
Movie S15 (.mov)
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