Figure 3

Phenotypic analysis of replacement clones. (A) Growth in HL-5 medium in shaken flasks. (B) Protein expression in vegetative cells; (i) Cell lysates prepared from equal numbers of cells were separated by SDS-PAGE, transferred onto nitrocellulose and immunoblotted as described in Materials and Methods. Blots were probed with Dictyostelium anti-Arp2 antibody. (ii) 20 μg of total protein was separated by SDS-PAGE, transferred onto PVDF and immunoblotted as described in Materials and Methods. Blots were probed with Dictyostelium anti-Arp2 and mAb 9E10 anti-myc antibodies. (C) All results are representative two to three experiments. Chemotaxis. Under-agar migratory response of cells to a gradient of folate. Arrows indicate the direction of cell migration (increasing folate concentration); (a) Scale bar is 50 μm. (a') Scale bar is 10 μm. Phototaxis. Response of cells spotted on nitrocellulose filters, to a lateral light source (48 hrs). The light source is always to the right of the image. Arrows indicate the direction of migration; (b) Scale bar is 5 mm. (b') Scale bar is 1 mm. Development. Fruiting body morphology of cells after development on nitrocellulose filters (48 hrs). (c) Scale bar is 1 mm. (c') Scale bar is 0.1 mm.