Journal: Frontiers in Immunology

Article Title: Male Macrophages and Fibroblasts from C57/BL6J Mice Are More Susceptible to Inflammatory Stimuli

doi: 10.3389/fimmu.2021.758767

Figure Lengend Snippet: Characterization of bone marrow derived macrophages (BMMs) and cardiac fibroblasts. (A) Flow cytometric analysis of the F4/80 and CD11b expression in BMMs. Representative images of BMMs stained against (B, C) F4/80 and (D, E) CD11b. Representative images of cardiac fibroblasts stained against (F, G) vimentin, (H, I) CD31 (endothelial marker; negative control) and (J, K) desmin (smooth muscle marker; negative control). Nuclei were stained with DAPI (C, E, G, I, K) Magnification: 40x; scale bar: 50 µm. Data are representative of 3 independent experiments with similar results.

Article Snippet: Fibroblasts were activated using 20 ng/ml TNF-α ( ) (PeproTech, Germany), 10 ng/ml TGF-β (PeproTech, Germany) ( ) or 10 ng/ml LPS (Sigma, Germany) for 24 h in fibroblast starvation medium (with 2.5% charcoal-stripped FCS, Biochrom, Germany).

Techniques: Derivative Assay, Expressing, Staining, Marker, Negative Control

Journal: Frontiers in Immunology

Article Title: Male Macrophages and Fibroblasts from C57/BL6J Mice Are More Susceptible to Inflammatory Stimuli

doi: 10.3389/fimmu.2021.758767

Figure Lengend Snippet: TNF-α, but not TGF-β, induces pro-fibrotic phenotype in mouse cardiac fibroblasts. Real-time PCR analyses of (A) MCP-1, (B) TGF-β, (C) IL-1β, and (D) Col1A1 performed with lysates from male and female mice cardiac fibroblasts treated with 20 ng/ml TNF-α for 24 h. Real-time PCR analyses of (E) MCP-1 and (F) Col1A1 performed with lysates from male and female mice cardiac fibroblasts treated with 10 ng/ml TGF-β for 24 h. Data are shown as means ± SEM (n = 6; independent experiments with technical duplicates). Data are normalized to the male untreated group. *p < 0.05, **p < 0.01, untreated vs. treated.

Article Snippet: Fibroblasts were activated using 20 ng/ml TNF-α ( ) (PeproTech, Germany), 10 ng/ml TGF-β (PeproTech, Germany) ( ) or 10 ng/ml LPS (Sigma, Germany) for 24 h in fibroblast starvation medium (with 2.5% charcoal-stripped FCS, Biochrom, Germany).

Techniques: Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: Male Macrophages and Fibroblasts from C57/BL6J Mice Are More Susceptible to Inflammatory Stimuli

doi: 10.3389/fimmu.2021.758767

Figure Lengend Snippet: Pro-inflammatory environment promotes a pro-inflammatory and pro-fibrotic fibroblast phenotype. Real-time PCR analyses of (A) MCP-1, (B) TNF-α, (C) NFκB, and (D) IL-1β performed with lysates from male and female mice cardiac fibroblasts cultivated with conditioned medium from M1 polarized BMMs for 24 h. Data are shown as means ± SEM (n = 6; independent experiments with technical duplicates). Data are normalized to the male untreated group. **p < 0.01, ***p < 0.001, untreated vs. treated; ## p < 0.01, male vs. female.

Article Snippet: Fibroblasts were activated using 20 ng/ml TNF-α ( ) (PeproTech, Germany), 10 ng/ml TGF-β (PeproTech, Germany) ( ) or 10 ng/ml LPS (Sigma, Germany) for 24 h in fibroblast starvation medium (with 2.5% charcoal-stripped FCS, Biochrom, Germany).

Techniques: Real-time Polymerase Chain Reaction