MshA pilus production correlates with c-di-GMP.

<p>Expression of the DGC VCA0956 was induced with varying amounts of IPTG. Intracellular c-di-GMP level was determined with LC-MS/MS and normalized to total protein in the extract (grey bars, right Y axis). Four independent samples were used for each induction condition. Oneway ANOVA followed by Dunnett’s Multiple Comparison Test determined significance. Surface MshA pilin was determined by ELISA (black bars, left Y axis). Three biological replicates were tested in triplicate. Results were normalized to MshA production in the WT strain. Surface MshA pili were significantly increased in all conditions compared to WT (Oneway ANOVA, Dunnett’s Multiple Comparison Test.) Also, there was a significant correlation between increased c-di-GMP and increased surface MshA pili (Pearson correlation, p = 0.0025, R² = 0.9199).