Journal: PLoS ONE

Article Title: The Role for Dickkopf-Homolog-1 in the Pathogenesis of Crohn’s Disease-Associated Fistulae

doi: 10.1371/journal.pone.0078882

Figure Lengend Snippet: (A) DKK-1 expression in control-transfected cells and after DKK-1-siRNA transfection with or without TGF-β (50 ng/ml) treatment (n = 5). (B) IL-13 (n = 4), (C) β6-Integrin (n = 5) and (D) SLUG mRNA expression (n = 4) in HT-29-IEC in control-transfected and transfected cells with or without application of TGF-β for 24 h. Asterisks indicate significant differences versus the respective control whereas # indicates significant differences versus TGF-β treatment and ° indicates significance in comparison to non-treated siRNA transfected cells. */# = p<0.05, **/## = p<0.01, ***/###/°°° = p<0.001.

Article Snippet: After starvation, HT-29 cells were basolaterally stimulated with either 100 ng/ml human recombinant Interleukin-13 (rhIL-13, R&D), 100 µg/ml of an anti-IL-13 antibody (provided by Novartis AG, Basel, Switzerland), 50 ng/ml human recombinant tumor necrosis factor alpha (TNF-α, Calbiochem/Merck, VWR), 50 ng/ml human recombinant tumor growth factor beta (TGF-β1, Calbiochem/Merck, VWR), 100 ng/ml muramyl dipeptide (MDP, InvivoGen) in a dimethyl sulfoxide (DMSO) – FuGene transfection reagent (Promega) solution, 100 ng/ml ultra-pure lipopolysaccharide (LPS, from E. coli, InvivoGen) and/or 24 µg/ml of an anti-TNF-antibody.

Techniques: Expressing, Transfection, Comparison

Journal: PLoS ONE

Article Title: The Role for Dickkopf-Homolog-1 in the Pathogenesis of Crohn’s Disease-Associated Fistulae

doi: 10.1371/journal.pone.0078882

Figure Lengend Snippet: (A) mRNA expression and (B) protein levels of β-Catenin (n = 2), mRNA levels of (C) DKK-1, (D) IL-13 and (E)TCF-4 in control and β-Catenin-knock-down HT-29 IEC stimulated with or without TGF-β (50 ng/ml) for 24 h (n = 5 each experiment).Asterisks indicate significant difference versus the no stimulated control cells whereas # indicates significant differences versus TGF-β treated control cells. */# = p<0.05, **/## = p<0.01, *** = p<0.001.

Article Snippet: After starvation, HT-29 cells were basolaterally stimulated with either 100 ng/ml human recombinant Interleukin-13 (rhIL-13, R&D), 100 µg/ml of an anti-IL-13 antibody (provided by Novartis AG, Basel, Switzerland), 50 ng/ml human recombinant tumor necrosis factor alpha (TNF-α, Calbiochem/Merck, VWR), 50 ng/ml human recombinant tumor growth factor beta (TGF-β1, Calbiochem/Merck, VWR), 100 ng/ml muramyl dipeptide (MDP, InvivoGen) in a dimethyl sulfoxide (DMSO) – FuGene transfection reagent (Promega) solution, 100 ng/ml ultra-pure lipopolysaccharide (LPS, from E. coli, InvivoGen) and/or 24 µg/ml of an anti-TNF-antibody.

Techniques: Expressing

Journal: PLoS ONE

Article Title: The Role for Dickkopf-Homolog-1 in the Pathogenesis of Crohn’s Disease-Associated Fistulae

doi: 10.1371/journal.pone.0078882

Figure Lengend Snippet: (A) DKK-1 mRNA expression in HT-29 cells stimulated with IL-13 (100 ng/ml) for 30 min, 24 h and 72 h (n = 3). (B) Effect on DKK-1 levels in HT-29-IEC treated for 24 h alone with IL-13, an anti-IL-13 antibody (100 µg/ml) or a combination of both (n = 12). (C) Expression pattern of DKK-1 in HT-29-spheroids exposed to IL-13 for 1, 5 and 7 days (n = 3). (D) DKK-1 mRNA expression in intestinal control fibroblasts (n = 5) and (E) in CD- patients with a fistulizing course (n = 4). Asterisks indicate significant differences versus the respective control whereas # indicates significant differences versus IL-13 treatment. */# = p<0.05, ## = p<0.01, ***/### = p<0.001.

Article Snippet: After starvation, HT-29 cells were basolaterally stimulated with either 100 ng/ml human recombinant Interleukin-13 (rhIL-13, R&D), 100 µg/ml of an anti-IL-13 antibody (provided by Novartis AG, Basel, Switzerland), 50 ng/ml human recombinant tumor necrosis factor alpha (TNF-α, Calbiochem/Merck, VWR), 50 ng/ml human recombinant tumor growth factor beta (TGF-β1, Calbiochem/Merck, VWR), 100 ng/ml muramyl dipeptide (MDP, InvivoGen) in a dimethyl sulfoxide (DMSO) – FuGene transfection reagent (Promega) solution, 100 ng/ml ultra-pure lipopolysaccharide (LPS, from E. coli, InvivoGen) and/or 24 µg/ml of an anti-TNF-antibody.

Techniques: Expressing

Journal: PLoS ONE

Article Title: The Role for Dickkopf-Homolog-1 in the Pathogenesis of Crohn’s Disease-Associated Fistulae

doi: 10.1371/journal.pone.0078882

Figure Lengend Snippet: Under chronic inflammatory conditions, TNF-α, and also IL-13, are present at high levels and exert inhibitory effects on DKK-1 mRNA expression in IEC. TNF-α further stimulates TGF-β secretion that promotes DKK-1 mRNA expression in IEC. The effects of the cytokine and the growth factor on DKK-1 are mediated via β-catenin/TCF. On a functional level, TGF-β induces IL-13 secretion via a β-catenin/DKK-1-dependent pathway. These mechanisms through synergism between TGF-β and IL-13 finally lead to an enhanced expression of genes associated with EMT and therefore to the formation of perianal fistulae associated to CD.

Article Snippet: After starvation, HT-29 cells were basolaterally stimulated with either 100 ng/ml human recombinant Interleukin-13 (rhIL-13, R&D), 100 µg/ml of an anti-IL-13 antibody (provided by Novartis AG, Basel, Switzerland), 50 ng/ml human recombinant tumor necrosis factor alpha (TNF-α, Calbiochem/Merck, VWR), 50 ng/ml human recombinant tumor growth factor beta (TGF-β1, Calbiochem/Merck, VWR), 100 ng/ml muramyl dipeptide (MDP, InvivoGen) in a dimethyl sulfoxide (DMSO) – FuGene transfection reagent (Promega) solution, 100 ng/ml ultra-pure lipopolysaccharide (LPS, from E. coli, InvivoGen) and/or 24 µg/ml of an anti-TNF-antibody.

Techniques: Expressing, Functional Assay