pro-inflammatory cytokine il-1β  (PeproTech)

Journal: Frontiers in Immunology

Article Title: The effects of inflammation on connexin 43 in chronic Chagas disease cardiomyopathy

doi: 10.3389/fimmu.2024.1440662

Figure Lengend Snippet: Morphological analysis and gene expression of pro-inflammatory cytokines in the hearts of uninfected and chagasic mice at 6 and 12 months after infection. (A, B) Representative micrographs of hematoxylin and eosin-stained heart sections of uninfected and chagasic mice at 12 months following infection. (C) Inflammatory cells quantified by morphometric analysis. (D, E) Micrographs of picrosirius red-stained heart sections of uninfected and chagasic mice. (F) Fibrotic area represented by percentage of collagen deposition in heart sections. Gene expression of pro-inflammatory cytokines (G) Tnf , (H) Il1b , (I) Ifng , and (J) Gja1 assessed by RT-qPCR using cDNA samples prepared from mRNA extracted from experimental mouse hearts. Values represent means ± S.E.M. of 5-6 mice per group. *** P < 0.001; ** P < 0.01; * P <0.05 compared to uninfected group.

Article Snippet: Following cell adherence to the coverslips, cultures were stimulated with a combination of pro-inflammatory cytokines (IL-1β, TNF, and IFN-γ; 10 ng/ml of each cytokine; Cell Signaling Technology) ( , ) in DMEM supplemented with 10% FBS for 48 h. Glass coverslips containing H9c2 cells were then washed with PBS and fixed in 4% paraformaldehyde (Thermo Scientific, Wilmington, DE) for 15 minutes at room temperature.

Techniques: Gene Expression, Infection, Staining, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: The effects of inflammation on connexin 43 in chronic Chagas disease cardiomyopathy

doi: 10.3389/fimmu.2024.1440662

Figure Lengend Snippet: Effect of inflammatory microenvironment on Cx43 distribution in vitro . (A) Experimental in vitro design involving H9c2 cells and iPSC-derived cardiomyocytes, with both cell types stimulated by pro-inflammatory cytokines (IL-1β, TNF, and IFN-γ, 10 ng/ml of each cytokine) for 48 hours for immunofluorescence analysis. (B) Immunofluorescence of H9c2 cells and iPSC-derived cardiomyocytes; (C) Analysis of Cx43 distribution by fluorescence intensity in H9c2 cells and iPSC-derived cardiomyocytes after 48 hours of stimulation with pro-inflammatory cytokines IL-1β, TNF, and IFN-γ. Cells were stained with WGA (red), cell nuclei with DAPI (blue), and total Cx43 with anti-Cx43 antibody (green). Images analyzed by confocal microscopy. Scale bars = 50 µm. WGA, Wheat Germ Agglutinin; Cx43, Connexin 43; IL-1β, Interleukin 1 beta; TNF, Tumor necrosis factor; IFN-γ, Interferon gamma; C, Cytoplasm; N, Cell Nucleus. Created with BioRender.com .

Article Snippet: Following cell adherence to the coverslips, cultures were stimulated with a combination of pro-inflammatory cytokines (IL-1β, TNF, and IFN-γ; 10 ng/ml of each cytokine; Cell Signaling Technology) ( , ) in DMEM supplemented with 10% FBS for 48 h. Glass coverslips containing H9c2 cells were then washed with PBS and fixed in 4% paraformaldehyde (Thermo Scientific, Wilmington, DE) for 15 minutes at room temperature.

Techniques: In Vitro, Derivative Assay, Immunofluorescence, Fluorescence, Staining, Confocal Microscopy

Journal: Frontiers in Immunology

Article Title: The effects of inflammation on connexin 43 in chronic Chagas disease cardiomyopathy

doi: 10.3389/fimmu.2024.1440662

Figure Lengend Snippet: Functional in vitro analysis of Cx43: Action Potential Duration and Lucifer Yellow dye transfer. (A) Schematic drawing of functional testing performed on iPSC-derived cardiomyocytes and H9c2 cardiomyocytes, stimulated with pro-inflammatory cytokines to respectively analyze the duration of action potential and dye transfer between neighboring cells. (B) After stimulation, action potential duration (APD) of iPSC-derived cardiomyocytes was determined at 10%, 50% and 90% levels. (C) Lucifer Yellow dye transfer test (green) performed in H9c2 cells (bright field) stimulated with pro-inflammatory cytokines IL-1β, TNF, and IFN-γ for 48 hours. Predominance of dye observed in cells at the margin where scalpel incision was made, while absence of stimulation resulted in dye reaching more distant cells; IL-1β = Interleukin 1 beta; TNF = Tumor necrosis factor; IFN-γ = Interferon gamma; CTRL = Control group; ***P< 0.001 compared to unstimulated cells. Scale bars = 100 μm.

Article Snippet: Following cell adherence to the coverslips, cultures were stimulated with a combination of pro-inflammatory cytokines (IL-1β, TNF, and IFN-γ; 10 ng/ml of each cytokine; Cell Signaling Technology) ( , ) in DMEM supplemented with 10% FBS for 48 h. Glass coverslips containing H9c2 cells were then washed with PBS and fixed in 4% paraformaldehyde (Thermo Scientific, Wilmington, DE) for 15 minutes at room temperature.

Techniques: Functional Assay, In Vitro, Derivative Assay, Control