FIGURE 5.

Anchorage of PKA is crucial for cell polarization and gradients of active PKA. Clone A cells were incubated with 5 μm stHt31P control peptide (A) or stHt31 (B) and then plated on laminin-1-coated coverslips and allowed to adhere and migrate in the presence of peptide. Cells and images were processed as described in Fig. 1. Graphics below the Fc/YFP images represent the average pixel intensity of the Fc/YFP ratio versus the distance in microns as displayed in Fig. 1. Bar graphics represent fluorescence intensity for YFP and Fc/YFP for regions noted by the red boxes. The bar in the left-hand Fc/YFP panel of A indicates 10 μm and is representative of the scale for all images. C, Clone A cells were incubated with 5 μm stHt31P control peptide or 5 μm stHt31 or left untreated and then plated onto laminin-1-coated 100-mm dishes for 50 min. Cells were then extracted, and active PKA was measured as described in Fig. 3. Representative data are shown.