Fig. 6. A comparison of ecmA induction and DimB nuclear import in Ax2 and cblA^– cells. (A) Ax2 and cblA^– cells, developed to the mound stage, were dissociated and induced with different concentrations of DIF in a monolayer assay. RNA was then extracted and ecmA concentration was evaluated by RT-PCR. (B) Mound cells were dissociated, suspended in KK2 buffer and treated with 100 nM DIF. At the indicated times, cells were fixed with 85% cold methanol and stained with anti-DimB antibody and then with Alexa-Fluor-488-conjugated secondary antibody (Molecular Probes). Samples were observed by confocal microscopy. This is a low affinity antibody. Hence the nuclear staining that is observed (white arrows) is relatively weak.