Fig. 7. GAP activity on AgRho1 proteins. (A) Comparison between GTP-locked Agrho1a and Agrho1b mutants with Aglrg1 and Agsac7. Spores from mycelia that are heterokaryotic for the indicated allele were spotted on medium selecting for the mutation in six-well plates. The plates were incubated at 30°C for 2 days. The speckles come from air bubbles in the agar plates. (B) AgSac7-GAP activity towards AgRho1 proteins. The activities were determined as described in Materials and Methods using 5 µg of each protein or buffer as a control. The optical density at 650 nm was determined against a control reaction with buffer only. (C) Localization of the AgRho1 GAPs. To exclude the possibility that the cytoplasmic signal of AgSAC7-GFP is only background fluorescence, we include a wild-type strain without GFP and performed western analysis using a GFP antibody (left bottom panel).