Figure 2

Dynacortin promotes cell polarization in response to cAMP. A, B. Western analysis shows that dynacortin is depleted to undetectable levels in an Ax2 and Ax3(Rep orf+) strains. Enlazin [42] is shown as a loading control. C, E. Wild-type control (carrying the empty vector) cells become highly polarized and migrate towards the needle, injecting 1 μM cAMP. D, F. Removal of dynacortin inhibits the ability of the cells to become highly polarized. The fraction of cells that do become polarized move towards the needle with wild-type kinetics (Table 1). Videos are representative of 4 to greater than 12 videos per genotype. Scale bar in C, 20 μm, applies to panels C-F. G. GFP-myoII localized at the rear cortex of the chemotaxing wild-type (myoII::GFP-myoII:pDRH; empty vector control) cell. H. In a polarized dynacortin-deficient cell (myoII::GFP-myoII:pDRH; dynhp:pLD1A15SN), GFP-myoII was partially enriched in the rear half of the cell. I. In a non-polarized dynacortin-deficient cell (myoII::GFP-myoII:pDRH; dynhp:pLD1A15SN), GFP-myoII did not enrich in the rear of the cell. G-I. Cells are migrating to the right of the panels. Scale bar in G is 10 μm and applies to G-I.