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Figure 3 | BMC Developmental Biology

Figure 3

From: Ectopic expression of cyclase associated protein CAP restores the streaming and aggregation defects of adenylyl cyclase a deficient Dictyostelium discoideumcells

Figure 3

Expression of GFP-CAP or GFP-N-pro-CAP restores aggregation defects but not complete development in aca- cells. (A) Phase contrast microscopy showing aggregate formation of starving AX2, AX2 expressing GFP-CAP (AX2-CAP), aca- cells expressing GFP-CAP (aca--CAP) or GFP-N-pro-CAP (aca--N-pro-CAP). 1 × 107 cells/ml resuspended in Soerensen phosphate buffer were starved in suspension and images were obtained with 20x bright field phase contrast microscope at indicated times (in hours, h). (B) aca- cells and cells expressing GFP-CAP or GFP-N-pro-CAP were plated on phosphate agar plates and allowed to undergo development and fruiting body formation. Images were obtained with 20x bright field phase contrast microscope at 20 h of development. The aca- cells remained as single cells whereas expression of GFP-CAP or GFP-N-pro-CAP in aca- cells led to formation of aggregation streams and mounds which however failed to proceed into late developmental stages and fruiting body formation in comparison to AX2 and AX2 expressing GFP-CAP. The larger aggregate size observed for AX2 expressing GFP-CAP is probably due to higher levels of contact site A protein as determined by western blot analysis (Figure 4A). The multicellular structure shown for AX2 is quite large. It might have resulted from a higher cell density in this area of the plate. Bars, 100 μm in both (A) and (B).

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