Figure 7. EIIA^Ntr production increased in response to 1,2-PDL and propionate.

(a and b) The transcriptional expression of ptsN was measured with or without 1,2-PDL (a) and propionate (b). β-Galactosidase activity from PptsN::lacZ was measured in Miller units (left), and the relative ptsN mRNA expression was determined using qRT-PCR (right). Each bar represents the average value of three independent experiments. (c and d) The stability of EIIA^Ntr-FLAG was evaluated by western blot analysis with or without 1,2-PDL (12.5 mM) (c) and propionate (10 mM) (d). At 5 h post inoculation, mRNA translation was halted by adding chloramphenicol (0.2 mg/ml), and that time was set as 0 min. Total proteins were harvested every 30 min, and equivalent amounts were subjected to SDS-PAGE analysis. The stability of EIIA^Ntr-FLAG was assessed using an anti-FLAG antibody. The levels of DnaK were used as controls. Full-length blots were presented in Supplementary Figure S6.