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Fig. 6. Rescue of culmination of P4H1- slugs by P4H1-expressing
cells, and accumulation of P4H1-expressing cells in the slug tip.
(A) Various ratios of parental (Ax3) and P4H1a- cells were
mixed at time 0, developed under ambient conditions, and harvested to count
detergent-resistant spores after 48 hours. The straight line describes the
calculated spore number if only Ax3 formed spores independent of an effect
from P4H1a- cells. The vertical bar indicates that 
20% of
cells are normally pre-stalk cells. (B) RFP-expressing normal Ax3 cells
were mixed with unlabeled cells, as indicated, in a 7:93 ratio at the start of
development. Flattened 16 hour slugs were photographed by phase contrast and
for RFP-fluorescence. (C) In the reverse experiment, RFP-expressing
P4H1- cells were mixed with unlabeled cells, as indicated, in a 3:7
ratio. Numbers below image represent the average ratio of mean intensity in
the pre-stalk region (demarcated by three white dots) to that of the pre-spore
(remaining) region, for multiple slugs (n=3-6). (D) Cultures
shown in panel B were allowed to culminate, and stalks were examined. Examples
in B and D are typical (n>10). (E) Spore numbers were
determined in the mixtures shown in D and for Ax3 and P4H1a- cells
alone (left panel). Percent of spores that were red in the mixtures is shown
in the right panel. Results are representative of two independent
experiments.
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