Figure 2

Analysis of YidC expression under stress. To monitor YidC production under stress, bacterial cultures at OD₆₀₀ of ~0.8 were exposed to different stress agents for 24 hours followed by immunoblotting of WCEs with anti-YidC antibodies. Differences in YidC levels between treated and untreated cultures were calculated after normalizing the band intensities of the anti-YidC blot with total protein in the corresponding lysates, as observed in the CBB-stained gel shown below the immunoblot (A). Band intensities were determined by densitometric scanning using the NIH ImageJ tool (B). Effect of cell surface stresses such as detergent (0.1% SDS) and heat (45 °C) on mRNA transcript of yidC was examined by qRT-PCR (C). Fold change in expression levels under each of these conditions was calculated after normalization with the levels of sigA in the same samples. Primers specific to whiB6 and sigB (Table 1) were used as positive control for heat and detergent stress, respectively. ND: not-determined. Blots represent three repeat experiments in (A). Bar graphs in (B) and (C) were obtained by plotting mean ± s.d. of three measurements. Asterisks indicate P ≤ 0.05, as determined by Student’s t test.