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. 2012 Sep 27;7(9):e46288. doi: 10.1371/journal.pone.0046288

Figure 4. Effect of rpoN R456A expression in TW14359ΔrpoN on σS stability, gadE and ler transcription.

Figure 4

(Panel A): Representative western immunoblots for σS in TW14359 (WT), TW14359ΔrpoN complemented with rpoN + (TW14359ΔrpoNpRAM-1), TW14359ΔrpoN (ΔrpoN), TW14359ΔrpoN complemented with rpoN R456A (TW14359ΔrpoNpRAM-2) before (t = 0 min) and 4 min after addition of tetracycline (Tet.). Stationary phase (Stat.) protein extracts were used as a positive control for σS, and TW14359ΔrpoS (ΔrpoS) as a negative control. Equal gel loading was controlled for by westerns for GroEL. (Panel B): Mean gadE and ler transcript levels by qRT-PCR are plotted against TW14359 (WT) and derivative strains from Panel A. Transcript levels are normalized to the 16S rRNA gene rrsH. Asterisks denote significant differences between WT and TW14359ΔrpoNpRAM-1 when compared to TW14359ΔrpoN and TW14359ΔrpoNpRAM-2 by Tukey’s HSD following a significant F-test (n≥3, p<0.05). Error bars indicate standard error of the mean.