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. 2011 Nov 7;6(11):e27148. doi: 10.1371/journal.pone.0027148

Figure 6. GTPase specificity is not altered in the mutant CDC-24 variants.

Figure 6

(A) Purification of mutant MBP-CDC24-GEFPH constructs used in in vitro GEF activity assays. A Coomassie stained SDS polyacrylamide gel loaded with equal volumes of eluate fractions of the indicated constructs is shown. Predicted fusion protein molecular weights (MW) are given below the corresponding lanes. (B) Mutant RhoGEF-PH fragments exhibit reduced in vitro GEF activity towards both RAC and CDC42 compared to the wild type construct irrespective of the temperature used for the assay. Nucleotide exchange activity is displayed normalized to the intrinsic exchange activity (“no GEF”) of each Rho GTPase (set to 100%). Data from two independent experiments (with two technical replicates of each sample) were performed for 21°C (blue columns); results are based on one experiment performed in technical triplicates; for 37°C (red columns). Error bars indicate standard deviation.