Journal: Veterinary immunology and immunopathology

Article Title: Epigenetic contribution to individual variation in response to lipopolysaccharide in bovine dermal fibroblasts

doi: 10.1016/j.vetimm.2013.10.015

Figure Lengend Snippet: Production of IL-8 by dermal fibroblasts following exposure to LPS (100 ng/ml) for 24 hours after undergoing pre-treatment with media alone, AZA, TSA, or AZA-TSA. Animals displaying differential responsiveness to LPS (Low responders and High responders; n=3/group) were investigated. In the absence of AZA and TSA, low and high responders showed differential responsiveness to LPS. However, following exposure to AZA and TSA either alone or in conjunction, differences in responsiveness to LPS were lost between the groups. In addition, exposure to AZA and TSA alone or in conjunction significantly increased IL-8 production in response to LPS. IL-8 production in the absence of LPS was not detectable. * indicated P<0.05. All values are mean ± S.D. (n=3/group).

Article Snippet: 2.3 Quantification of IL-8 The concentration of IL-8 in fibroblast conditioned media was quantified by a custom sandwich ELISA previously described ( Kandasamy et al., 2011 ) using mouse anti-bovine IL-8 monoclonal antibody (clone 170.13 generously gifted by S. Maheswaren, University of Minnesota) and biotinylated goat anti-human IL-8 antibody (R&D Systems Inc., Minneapolis, MN) as capture and detection antibodies, respectively, and recombinant bovine IL-8 (Thermo Scientific, Rockford, IL) as assay standard.

Techniques:

Journal: Veterinary immunology and immunopathology

Article Title: Epigenetic contribution to individual variation in response to lipopolysaccharide in bovine dermal fibroblasts

doi: 10.1016/j.vetimm.2013.10.015

Figure Lengend Snippet: Comparison of gene expression following exposure to LPS at hour 0 for low and high responding cultures under both control (left panel) and epigenetic modification (right panel) conditions. IL-8 (A and B), IL-6 (C and D), and TNF-α (E and F) mRNA were all targeted. Values are expressed as dCt using β-actin expression as the endogenous control. All values are mean ± S.D. (n=3/group). * indicates significantly different expression levels (P < 0.05).

Article Snippet: 2.3 Quantification of IL-8 The concentration of IL-8 in fibroblast conditioned media was quantified by a custom sandwich ELISA previously described ( Kandasamy et al., 2011 ) using mouse anti-bovine IL-8 monoclonal antibody (clone 170.13 generously gifted by S. Maheswaren, University of Minnesota) and biotinylated goat anti-human IL-8 antibody (R&D Systems Inc., Minneapolis, MN) as capture and detection antibodies, respectively, and recombinant bovine IL-8 (Thermo Scientific, Rockford, IL) as assay standard.

Techniques: Expressing, Modification

Journal: Veterinary immunology and immunopathology

Article Title: Epigenetic contribution to individual variation in response to lipopolysaccharide in bovine dermal fibroblasts

doi: 10.1016/j.vetimm.2013.10.015

Figure Lengend Snippet: Expression of IL-8 (A), IL-6 (B), TNF-α (C), CCL20 (D), TLR4 (E) and SAA3 (F) mRNA following exposure to LPS for three low responding cultures. Expression was analyzed for response under both control and epigenetic treatment conditions. * indicates significantly different (P < 0.05) gene expression due to epigenetic modification. Values are expressed as dCt using β-actin expression as the endogenous control. All values are mean ± S.D.

Article Snippet: 2.3 Quantification of IL-8 The concentration of IL-8 in fibroblast conditioned media was quantified by a custom sandwich ELISA previously described ( Kandasamy et al., 2011 ) using mouse anti-bovine IL-8 monoclonal antibody (clone 170.13 generously gifted by S. Maheswaren, University of Minnesota) and biotinylated goat anti-human IL-8 antibody (R&D Systems Inc., Minneapolis, MN) as capture and detection antibodies, respectively, and recombinant bovine IL-8 (Thermo Scientific, Rockford, IL) as assay standard.

Techniques: Expressing, Modification