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Results: 7

1.
FIG 7 

FIG 7 . From: Haemophilus ducreyi Hfq Contributes to Virulence Gene Regulation as Cells Enter Stationary Phase.

Serum bactericidal assays. (A) Percent survival of 35000HP, 35000HP∆hfq, and the dsrA mutant in 50% NHS, calculated as follows: (geometric mean CFU in NHS/geometric mean CFU in heat-inactivated NHS) × 100. (B) Percent survival of 35000HP(pACYC177), 35000HP∆hfq(pACYC177), 35000HP∆hfq(pML129), and 35000HP∆dsrA(pACYC177) in 50% NHS, calculated as follows: (geometric mean CFU in NHS/geometric mean CFU in heat-inactivated NHS) × 100. Values are means ± SD of the results of 5 independent experiments.

Dharanesh Gangaiah, et al. mBio. 2014 Jan-Feb;5(1):e01081-13.
2.
FIG 1 

FIG 1 . From: Haemophilus ducreyi Hfq Contributes to Virulence Gene Regulation as Cells Enter Stationary Phase.

Venn diagram showing the overlap in differential gene expression across different growth phases. The up- and downregulated genes are indicated by up (↑) and down (↓) arrows, respectively. The total number of differentially regulated genes in different comparisons is indicated in bold outside the Venn diagram. Stationary/mid-log, genes differentially expressed in stationary phase relative to mid-log phase; Stationary/transition, genes differentially expressed in stationary phase relative to transition phase; Transition/mid-log, genes differentially expressed in transition phase relative to mid-log phase.

Dharanesh Gangaiah, et al. mBio. 2014 Jan-Feb;5(1):e01081-13.
3.
FIG 2 

FIG 2 . From: Haemophilus ducreyi Hfq Contributes to Virulence Gene Regulation as Cells Enter Stationary Phase.

qRT-PCR validation of growth-phase-dependent differences in gene expression derived from RNA-Seq. (A) The fold change in the expression of target genes in stationary phase relative to mid-log phase. The expression levels of target genes were normalized to that of dnaE. The data represent the means ± SD of the results of four independent experiments. (B) Correlation between the fold changes obtained from qRT-PCR and RNA-Seq analysis. The diagonal line represents the power trendline (R2 = 0.82).

Dharanesh Gangaiah, et al. mBio. 2014 Jan-Feb;5(1):e01081-13.
4.
FIG 4 

FIG 4 . From: Haemophilus ducreyi Hfq Contributes to Virulence Gene Regulation as Cells Enter Stationary Phase.

Comparison of the expression patterns of the genes altered by inactivation of hfq to those of the genes differentially expressed in stationary phase relative to mid-log phase. (A) Venn diagram showing the overlap in the genes altered by inactivation of hfq and the genes differentially expressed in stationary phase relative to mid-log phase. The total number of genes in each comparison is indicated in bold outside the Venn diagram. Potential Hfq targets, genes differentially expressed in 35000HPΔhfq::cat relative to 35000HP; Stationary/mid-log, genes differentially expressed in stationary phase relative to mid-log phase. The significance of the overlap was tested using the chi-square test (P < 2.2e-16). (B) Inverse correlation between the fold changes of the genes altered by inactivation of hfq and those of the genes differentially expressed in stationary phase relative to mid-log phase. Only genes that overlapped in their expression patterns as shown in panel A were used for the correlation analysis.

Dharanesh Gangaiah, et al. mBio. 2014 Jan-Feb;5(1):e01081-13.
5.
FIG 3 

FIG 3 . From: Haemophilus ducreyi Hfq Contributes to Virulence Gene Regulation as Cells Enter Stationary Phase.

Relative expression levels of selected H. ducreyi genes in 35000HPhfq::cat. (A) Expression levels of 15 selected genes in 35000HPhfq::cat compared to 35000HP were measured by DNA microarray (black bars) or real-time RT-PCR (white bars) as described in Materials and Methods. These data are from a representative experiment. (B) Correlation between log2 values obtained by DNA microarray analysis and qRT-PCR analysis. The diagonal line represents the power trendline (R2 = 0.8760).

Dharanesh Gangaiah, et al. mBio. 2014 Jan-Feb;5(1):e01081-13.
6.
FIG 6 

FIG 6 . From: Haemophilus ducreyi Hfq Contributes to Virulence Gene Regulation as Cells Enter Stationary Phase.

H. ducreyi microcolony formation assay. The relative abilities of 35000HP, 35000HPhfq::cat, 35000HPhfq::cat(pML129), and 35000HPhfq::cat(pACYC177) to form microcolonies was tested by incubating these strains with Hs27 fibroblasts. The larger pictures were taken at a ×14 magnification. The contents of the white boxes are shown to the right of each picture at a ×40 magnification. Results of a representative experiment are shown. Note that the hfq mutant formed bacterial cell aggregates but that these clusters of bacterial cells were much less compact than those formed by the parent and complemented strains.

Dharanesh Gangaiah, et al. mBio. 2014 Jan-Feb;5(1):e01081-13.
7.
FIG 5 

FIG 5 . From: Haemophilus ducreyi Hfq Contributes to Virulence Gene Regulation as Cells Enter Stationary Phase.

Inactivation of the H. ducreyi hfq gene has a positive effect on the synthesis of known virulence determinants. Data represent the results of Western blot analysis of whole-cell lysates from 35000HP (lane 1), 35000HPhfq::cat (lane 2), 35000HPhfq::cat(pML129) (lane 3), and 35000HPhfq::cat(pACYC177) (lane 4) probed with LspA1 MAb 40A4, LspA2 MAb 1H9, an LspB polyclonal antibody, a DsrA polyclonal antibody, or a Flp1 polyclonal antibody. The LspA1 and LspA2 proteins do not have discrete banding patterns in Western blots but instead form smears (27). PAL MAb 3B9 was used to confirm equivalent loading among lanes.

Dharanesh Gangaiah, et al. mBio. 2014 Jan-Feb;5(1):e01081-13.

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