Figure 7
From: Glucose Levels Alter the Mga Virulence Regulon in the Group A Streptococcus
Mga is differentially phosphorylated based on glucose availability. The isogenic M4 mga mutant strain KSM547 complemented in trans with the His6- tagged WT strain (pKSM808) or phosphoablative A/A/A mga mutant (pKSM871) was grown in THY or C medium to the late exponential growth phase. Shown is a representative image of multiple replicates. (A) Equal concentrations of cell lysates of WT and A/A/A expressing strains grown in each media were resolved on a Phos-tag gel and immunoblotted using polyclonal αMga4 antibody. Bands corresponding to non-phosphorylated Mga and phosphorylated Mga are indicated by arrows. (B) Mga levels in same cell lysates were determined by 10% SDS-PAGE and immunoblotting using polyclonal αMga4 antibody. (B) Densitometric ratio of WT Mga~P-His6 bands to total Mga-His6 from THY and (C) Media lysates after separation on Phos-tag and Western blotting with a-Mga. Results were normalized to the same ratio in the respective A/A/A Mga-His6 control lanes from 2 biological replicates. Standard Error of the Mean (SEM) is indicated.
