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Requirements for the adenylyl cyclases in the development of Dictyostelium

Christophe Anjard, Fredrik Söderbom* and William F. Loomis{ddagger}

Center for Molecular Genetics, Division of Biology, University of California San Diego, La Jolla, CA 92093, USA
* Present address: Dept. of Microbiology, Swedish University of Agricultural Sciences Box 7025, 750 07 Uppsala, Sweden



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  		Fig. 1. Proportion of visible and viable spores. Cells were developed on filters for 18 to 24 hours and collected to determine the percentage of visible (stippled bars) and viable (hatched bars) spores compared to the total number of cells. The times at which the maximal proportion of viable spores was found were: 22 hours for acrA- cells of strain DG1100; 18 hours for acrA-/K cells; 20 hours for acrA-/ACA* cells; 24 hours for acrA+ cells of strain AX4. Each experiment was repeated at least 3 times.

 


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  		Fig. 2. Terminal structures of strains AK631 and TL130. Cells were deposited on filters at high density and allowed to develop for up to 36 hours before being photographed. As previously described (Wang and Kuspa, 1997) the acaA- PKA-Cover cells of strain AK631 form fairly normal fruiting bodies. Most of the cells of strain TL130 (acaA- acrA- PKA-Cover) were found in tipped mounds but a few were seen to form small finger-like structures.

 


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  		Fig. 3. Expression of the cell-type-specific genes ecmA and cotB. Total RNA was collected from strains AX4 (wild type), AK631 (acaA- PKA-Cover) and TL130 (acaA- acrA- PKA-Cover) after 20 hours of development, separated on agarose gels and transferred to nylon membranes. Northern blots were sequentially probed with the prestalk-specific gene ecmA and the prespore-specific gene cotB.

 


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  		Fig. 4. Calcofluor staining. Cells of strains AX4 and TL130 were developed on filters for 21 hours (A-D,F) or 24 hours (E) before being stained with 0.1% Calcofluor White ST and observed under a fluorescence microscope. Wild-type AX4 cells (A,C,E); acaA-, acrA- PKA-Cover cells of strain TL130 (B,D,F). Scale bars, 5 µm.

 

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© The Company of Biologists Ltd 2001