Journal: PLoS ONE
Article Title: During bacteremia, Pseudomonas aeruginosa PAO1 adapts by altering the expression of numerous virulence genes including those involved in quorum sensing
doi: 10.1371/journal.pone.0240351
Figure Lengend Snippet: Fold change in gene expression at each time point is relative to its expression in LBB at the same time point (value of 1, dotted line). Dashed gray line indicates the cut-off for significance, fold change ≥ 2.00. (A and B) Fold change in expression at 4 h of growth post-inoculation of 18 genes that regulate the QS systems (expression in WBHVs compared to that in LBB). Values represent the means of 3 replicate tests from 3 independent samples (WBHV); the bar represents the median. The data shown are based on the complete RNA-Seq analysis; black asterisk * over a black bar indicates that change in gene expression was considered significant among all 3 individual HV samples using fold change ≥ 2.00 and q values ≤ 0.05; green asterisks indicate significant difference in gene expression for that individual sample. Values in shades of blue, all or 2 of 3 values for gene expression were above 0 among the three samples; values in shades of purple, some values for gene expression were above 0 and some values were below 0 (A); values in shades of red, all or 2 of 3 values for gene expression were 0 or below among the three samples (B). (C) Comparison of the effect of WBHVs with that of 10% PHS on the expression of selected QS regulatory genes at 4 h of growth of PAO1 post-inoculation. RNA-Seq analysis of the effect of WBHV is described in (A). To determine the effect of 10% PHS, PAO1 was grown in LBB or LBBS and the expression level was determined by qRT-PCR. Values represent the means of replicate tests from 3 independent samples; the bar represents the median. Significance was determined by unpaired two-tailed t test. Black asterisk over black bar, WBHV:LBBS; dark red asterisk, WBHV:LBB; dark blue asterisk, LBBS:LBB; *, P <0.05, **, P <0.01. (D) The effect of 10% PHS on the expression of selected QS regulatory genes in PAO1 grown in either LBB or LBBS for 4 h and 8 h post-inoculation. The level of expression of the indicated genes was determined using qRT-PCR. Values represent the means of 3 independent experiments ± SEM. Significance was determined by unpaired two-tailed t test; *, P <0.05; **, P < 0.01; ***, P < 0.001; ****, P <0.0001; LBBS:LBB at 4 h, blue *; LBBS:LBB at 8 h, red *; LBBS-4h:LBBS-8h, black *.
Article Snippet: For experiments involving pooled normal human serum (PHS) (MilliporeSigma, St. Louis, MO), cultures were grown in either LBB or LBB supplemented with 10% human serum (LBBS) under conditions described above for LBB, and samples were collected at specific time points within the growth cycle.
Techniques: Gene Expression, Expressing, RNA Sequencing, Comparison, Quantitative RT-PCR, Two Tailed Test
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