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Open Access Highly Accessed Research article

On the effects of cycloheximide on cell motility and polarisation in Dictyostelium discoideum

Margaret Clotworthy* and David Traynor

Author Affiliations

MRC Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, England

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BMC Cell Biology 2006, 7:5  doi:10.1186/1471-2121-7-5

Published: 24 January 2006

Additional files

Additional File 1:

Comparison of the effects of cycloheximide on fluid phase uptake, as measured using FITC-dextran, in the wild-type strain Ax2 and a cycloheximide resistant mutant, HH31. Fluid phase uptake is inhibited by 2 mM cycloheximide in Ax2 but not in the cycloheximide-resistant mutant, HH31. Amoebae were incubated in various concentrations of cycloheximide for 30 minutes, followed by the addition of 2 mM FITC-dextran. The data for each experiment were normalised with respect to the 30 minute time point for Ax2 in the absence of cycloheximide. Movies: All movies were taken as sections every 2 seconds, using a 60× (1.4 NA) oil immersion lens on a Nikon Eclipse TE300 microscope fitted with the BioRad Radiance confocal system. The movies are played at 6 frames per second. The movies were generated by merging the DIC (grey) and fluorescence images. The fluorescence intensity is graded from red (low intensity) to white (high intensity) for 3 of the movies, ABP120 with and without cycloheximide, and actin without cycloheximide. For the other movies, the fluorescence is in green and the relative intensity is not indicated.

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Open Data

Additional File 2:

ABP120 with cycloheximide. Aggregation-competent amoebae were allowed to settle on a chambered coverglass for 10 minutes before being incubated with 2 mM cycloheximide for 1 hour and then stimulated with a micropipette filled with 1 μM cAMP in the presence of cycloheximide.

Format: MP4 Size: 2.6MB Download file

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Open Data

Additional File 3:

ABP120 without cycloheximide. Aggregation-competent amoebae were allowed to settle on a chambered coverglass for 10 minutes before being stimulated with a micropipette filled with 1 μM cAMP in the presence of cycloheximide.

Format: MP4 Size: 4.2MB Download file

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Open Data

Additional File 4:

Actin with cycloheximide. Aggregation-competent amoebae were allowed to settle on a chambered coverglass for 10 minutes before being incubated with 2 mM cycloheximide for 1 hour and then stimulated with a micropipette filled with 1 μM cAMP in the presence of cycloheximide.

Format: MOV Size: 2.9MB Download file

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Open Data

Additional File 5:

Actin without cycloheximide. Aggregation-competent amoebae were allowed to settle on a chambered coverglass for 10 minutes before being stimulated with a micropipette filled with 1 μM cAMP in the presence of cycloheximide.

Format: MP4 Size: 3.7MB Download file

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Open Data

Additional File 6:

CRAC with cycloheximide. Aggregation-competent amoebae were allowed to settle on a chambered coverglass for 10 minutes before being incubated with 2 mM cycloheximide for 1 hour and then stimulated with a micropipette filled with 1 μM cAMP in the presence of cycloheximide.

Format: MOV Size: 3.1MB Download file

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Open Data

Additional File 7:

CRAC without cycloheximide. Aggregation-competent amoebae were allowed to settle on a chambered coverglass for 10 minutes before being stimulated with a micropipette filled with 1 μM cAMP in the presence of cycloheximide.

Format: MOV Size: 2.5MB Download file

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Open Data

Additional File 8:

PI3K with cycloheximide. Aggregation-competent amoebae were allowed to settle on a chambered coverglass for 10 minutes before being incubated with 2 mM cycloheximide for 1 hour and then stimulated with a micropipette filled with 1 μM cAMP in the presence of cycloheximide.

Format: MOV Size: 4.2MB Download file

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Open Data

Additional File 9:

PI3K without cycloheximide. Aggregation-competent amoebae were allowed to settle on a chambered coverglass for 10 minutes before being stimulated with a micropipette filled with 1 μM cAMP in the presence of cycloheximide.

Format: MOV Size: 9.7MB Download file

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Open Data