Figure 2.

 Synthesis of DIF-1 during differentiation of wild-type and car4⁻ cells. Wild-type JH10 and car4 null cells were differentiated in suspension culture with 1 μCi/ml of ³⁶Cl. For the first 5 hr, cAMP was added at 6-min intervals to a final concentration of 30 nm. Subsequently, cultures were left untreated or adjusted to 300 μm cAMP. Cells and media were extracted twice with 1 volume of hexane, and extracts were pooled, dried, resuspended in ETOH, and analyzed by thin layer chromatography. Radiolabeled DIF-1 was identified by comigration with purified DIF-1. Quantification by PhosphorImager analyses yielded differences in DIF-1 accumulation of ∼50% between the two strains.