QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 4. Prespore and spore differentiation in adenylyl cyclase mutants. (A) Slugs of wild-type, acrA-, acg- and acrA-/A15::ACGcat cells that had migrated for 2-3 hours were dissociated and stained with a spore-specific antiserum and GARFITC. Cell nuclei were counterstained with DAPI. The percentage of prespore cells (cells with at least 3-4 fluorescent vacuoles) relative to DAPI-stained cells was determined. Means and SE of four experiments are presented. Significant differences (P>0.95) between datasets connected by brackets, as determined by Kruskal-Wallis ANOVA on ranks using SigmaStat software (Systat, San Jose, US), are indicated by asterisks. (B) Wild-type, acrA-, acg- and acrA-/A15::ACGcat cells were developed on PB agar until fruiting bodies had formed. Total RNA was extracted at 2-hour intervals, size-fractionated on 1.5% agarose gels containing 2.2 M formaldehyde and transferred to nylon membranes (Nellen et al., 1987). The four northern blots were hybridized in the same batch to a [³²P]dATP-labelled CotB probe at 65^oC, then stripped and reprobed with the constitutively expressed gene 1G7 (Williams et al., 1987). (C) Three-day-old fruiting bodies of acrA-, acg- and acrA-/A15::ACGcat cells were transferred to a glass slide and stained with the cellulose dye Calcofluor at 0.03% (w/v) final concentration. The preparations were photographed under UV by fluorescence microscopy to visualize the Calcofluor-stained spores in the presence of a low level of transillumination to obtain a phase contrast image of the remaining amoebae. Scale bar: 10 µm.

Return to article