Fig. 4. Effects of cadA gene inactivation on cell differentiation. Equal numbers of KAX3 cells and cadA^– cells were deposited on 2% non-nutrient agar for development. (A) Reduction in spore yields in cadA^– strains. Fruiting bodies were collected from the agar dish after 2 days of development and the total number of spores was estimated. The relative yield of spores was calculated. Data represent mean±s.d. of three experiments. (B) Increase in Neutral Red-strained cells in cadA^– slugs. KAX3 (wild-type) () and cadA-10 cells () were labeled with Neutral Red and then deposited on 2% non-nutrient agar plates at 5x10⁵ cells/cm² for development in the dark at 22°C. Cells at different stages of development were collected and dissociated in 17 mM phosphate buffer (pH 6.4), containing 20 mM EDTA, and the percentage of Neutral Red-stained cells was determined. Data represent mean±s.d. of three experiments.