Results: 3

1.
Figure 2

Figure 2. STAT3 and RelA in hepatocytes are required for the liver transcriptional response to pneumonia.. From: Hepatocyte-specific mutation of both NF-?B RelA and STAT3 abrogates the acute phase response in mice .

(A) Heat map indicates relative gene expression for all 1,157 transcripts significantly (FDR < 0.05) changed in control mouse livers following 24 hours of pneumococcal pneumonia (serotype 3; 106 CFU). Each column represents an individual control (–) or mutant (+) mouse, and each row is a distinct transcript. Red or green shows increased or decreased expression, respectively. (B) The effect of STAT3 and RelA deletion on pneumonia-influenced genes is indicated, using FDR < 0.05 to differentiate transcripts that are: (a) completely dependent (significantly changed during pneumonia only in CRE mice); (b) partially dependent (changed during pneumonia in both genotypes, but significantly less so in CRE+ mice); and (c) independent (changed during pneumonia in both genotypes, with no significant difference between genotypes).

Lee J. Quinton, et al. J Clin Invest. 2012 May 1;122(5):1758-1763.
2.
Figure 1

Figure 1. Deletion of STAT3 and RelA in hepatocytes eliminates acute phase protein induction.. From: Hepatocyte-specific mutation of both NF-?B RelA and STAT3 abrogates the acute phase response in mice .

(A) PCR confirmed gene incorporations of Cre-recombinase driven by an albumin promoter (Alb-Cretg) and LoxP insertions in Rela and Stat3 alleles. TCR-δ was an amplification control. (B) Immunoblots revealed liver-specific deletion of STAT3 and RelA in CRE+ mice. (C) qRT-PCR was performed to determine Saa1, Sap, and Lbp mRNA induction in response to intratracheal (i.t.) S. pneumoniae (serotype 3; 106 CFU), i.t. E. coli (106 CFU), i.v. cytokines (TNF-α, IL-1β, and IL-6), or s.c. casein. Values represent fold induction compared with that in uninfected CRE mice, expressed as geometric means ± geometric SEM. *P < 0.05 (n = 3–10 mice/group). (D) Plasma concentrations of SAA and SAP were quantified in samples collected 24 hours after i.t. S. pneumoniae using ELISA and expressed as mean ± SEM (n = 3–11 mice/group). *P < 0.05, effect of infection; P < 0.05, effect of genotype.

Lee J. Quinton, et al. J Clin Invest. 2012 May 1;122(5):1758-1763.
3.
Figure 3

Figure 3. Liver STAT3 and RelA are required for blood-borne host defense during pneumonia.. From: Hepatocyte-specific mutation of both NF-?B RelA and STAT3 abrogates the acute phase response in mice .

(A) Living bacteria were enumerated in blood collected 48 hours after intratracheal S. pneumoniae serotype 3 (104 CFU). Data points represent individual mice, and lines indicate medians (n = 16–18). (B) Survival through 48 hours was documented after intratracheal instillation of S. pneumoniae serotype 19F (106 CFU) (n = 15). (C and D) Effects of serum on opsonophagocytosis were measured by quantifying fluorescence/cell using flow cytometry after J774A.1 mouse macrophage-like cells were incubated with fluorescent S. pneumoniae and mouse serum. Representative histograms and the percentage of cells fluorescent illustrate effects of (C) pneumonia (in control CRE- mice) or (D) genotype (during pneumonia). Results represent 3 experiments, each containing pooled sera from distinct mice. (E) C3 deposition was measured on serum-opsonized S. pneumoniae by flow cytometry. Data represent mean ± SEM of the percentage of C3+ bacteria exposed to serum from different mice (n = 4–7). Colors in CE correspond to serum collected from uninfected mice (green), 24-hour infected CRE control mice (red), or 24-hour infected CRE+ mutant mice (blue). Shaded curves represent background (nonopsonized) bacterial fluorescence (E) or background J774A.1 fluorescence (C and D), which was similar for cells alone, cells exposed to nonopsonized bacteria, or experiments conducted at 0°C. *P < 0.05, difference between groups.

Lee J. Quinton, et al. J Clin Invest. 2012 May 1;122(5):1758-1763.

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