Figure 5

D-site mutations were correlated with repressed midgut ERK phosphorylation levels in A. gambiae compared to mosquitoes transformed with catalytically active MEK. A, Representative western blot of ERK phosphorylation in midgut epithelia from transformed mosquitoes examined at 2 h after blood feeding. Values below the blots indicate relative fold change in pERK levels in the midgut for each group of A. gambiae. Phosphorylated ERK levels were normalized to GAPDH and then to pERK levels in wtMEK, which is set at 1.00. B, Fold changes in ERK phosphorylation levels are indicated as trendlines between samples from matched treatment groups for pMEK2 and pMEK5 overexpression. Note that in one replicate, relative values of phosphorylation for pMEK2 and pMEK5 transformed A. stephensi were below the level of controls (e.g., < 1), but nonetheless revealed the same trend of decreasing phosphorylation. A Pearson’s r test was performed to assess the relationship between docking site mutations and ERK phosphorylation levels in midgut tissue of transformed F0 females (r = 0.7799, P = 0.06).