Journal: Frontiers in Cardiovascular Medicine

Article Title: Sex- and age-related differences in the inflammatory properties of cardiac fibroblasts: impact on the cardiosplenic axis and cardiac fibrosis

doi: 10.3389/fcvm.2023.1117419

Figure Lengend Snippet: Sex- and age-dependent differences in left ventricular macrophage presence. To determine LV presence of CD68 + macrophages ( A ), including the anti-inflammatory CD68 + CD206 + ( B ) and pro-inflammatory CD68 + CD206 − ( C ) subsets, immunofluorescence on tissue sections was performed. After the respective stainings, pictures were acquired at 20x magnification and macrophage populations were manually counted per Field of View (FoV). Amount of CD68 + cells were expressed as number of cells/FoV. CD68 + CD206 + and CD68 + CD206 − subpopulations were quantified as percentage (%) of CD68 + cells. Representative images [( D ); 40x magnification; scale bar = 50 μ m] illustrate the different stainings on LV cryosections of male and female mice at the age of 3- and 12-months. Data are depicted as scatter plots with bars (male mice: gray bars; female mice: wine red bars), showing individual data points and the corresponding mean ± SEM. For statistical analysis, Two-way ANOVA with Fisheŕs LSD post hoc test was performed ( § p < 0.05, §§ p < 0.01, §§§ p < 0.001, §§§§ p < 0.0001 male vs. female mice; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. 3-months old male mice; with n = 7 for 3-months old male mice, n = 9 for 3-months old female mice, n = 8 for 12-months old male mice, n = 9 for 12-months old female mice).

Article Snippet: Acetone-fixed and -permeabilized LV transverse OCT sections, blocked with 1% bovine serum albumin in PBS, were stained with mouse anti-α-actinin (A7811, Sigma-Aldrich; secondary antibody: FITC donkey anti-mouse, 715-096-150, JacksonImmunoResearch, West Grove, PA, USA), rat anti-mouse CD68 (MCA 1957GA, Bio-Rad; Hercules, CA, USA; secondary antibody: Alexa Fluor 647 Donkey anti-rat IgG, 712-605-153, JacksonImmunoResearch) and goat anti-mouse CD206 (AF2535, R&D, Wiesbaden, Germany; secondary antibody: Rhodamine Red-X Donkey anti-goat, 705-295-147, JacksonImmunoResearch).

Techniques: Immunofluorescence

Journal: Frontiers in Cardiovascular Medicine

Article Title: Sex- and age-related differences in the inflammatory properties of cardiac fibroblasts: impact on the cardiosplenic axis and cardiac fibrosis

doi: 10.3389/fcvm.2023.1117419

Figure Lengend Snippet: Sex- and age-dependent differences in left ventricular macrophage presence. To determine LV presence of CD68 + macrophages ( A ), including the anti-inflammatory CD68 + CD206 + ( B ) and pro-inflammatory CD68 + CD206 − ( C ) subsets, immunofluorescence on tissue sections was performed. After the respective stainings, pictures were acquired at 20x magnification and macrophage populations were manually counted per Field of View (FoV). Amount of CD68 + cells were expressed as number of cells/FoV. CD68 + CD206 + and CD68 + CD206 − subpopulations were quantified as percentage (%) of CD68 + cells. Representative images [( D ); 40x magnification; scale bar = 50 μ m] illustrate the different stainings on LV cryosections of male and female mice at the age of 3- and 12-months. Data are depicted as scatter plots with bars (male mice: gray bars; female mice: wine red bars), showing individual data points and the corresponding mean ± SEM. For statistical analysis, Two-way ANOVA with Fisheŕs LSD post hoc test was performed ( § p < 0.05, §§ p < 0.01, §§§ p < 0.001, §§§§ p < 0.0001 male vs. female mice; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. 3-months old male mice; with n = 7 for 3-months old male mice, n = 9 for 3-months old female mice, n = 8 for 12-months old male mice, n = 9 for 12-months old female mice).

Article Snippet: Acetone-fixed and -permeabilized LV transverse OCT sections, blocked with 1% bovine serum albumin in PBS, were stained with mouse anti-α-actinin (A7811, Sigma-Aldrich; secondary antibody: FITC donkey anti-mouse, 715-096-150, JacksonImmunoResearch, West Grove, PA, USA), rat anti-mouse CD68 (MCA 1957GA, Bio-Rad; Hercules, CA, USA; secondary antibody: Alexa Fluor 647 Donkey anti-rat IgG, 712-605-153, JacksonImmunoResearch) and goat anti-mouse CD206 (AF2535, R&D, Wiesbaden, Germany; secondary antibody: Rhodamine Red-X Donkey anti-goat, 705-295-147, JacksonImmunoResearch).

Techniques: Immunofluorescence

Journal: Frontiers in Cardiovascular Medicine

Article Title: Sex- and age-related differences in the inflammatory properties of cardiac fibroblasts: impact on the cardiosplenic axis and cardiac fibrosis

doi: 10.3389/fcvm.2023.1117419

Figure Lengend Snippet: Graphical abstract representing the impact of cardiac fibroblasts as central players in sex- and age-related differences in cardiac fibrosis and inflammation. The first panel of the figure illustrates the higher inflammatory potential of male cFB vs. female cFB at the age of 3-months. Analysis of the spleen revealed a higher number of anti-inflammatory Ly6C low monocytes and a lower number of pro-inflammatory Ly6C high monocytes in the spleen of male vs. female animals, supporting the hypothesis that due to the higher inflammatory potential of male cFB, the pro-inflammatory Ly6C high monocytes migrated towards the heart. In contrast, cFB derived from female mice have a lower inflammatory potential mediating migration of pro-inflammatory Ly6C high monocytes towards the heart, as reflected by a higher number of pro-inflammatory Ly6C high monocytes in the spleen of female vs. male animals. In agreement, the percentage of pro-inflammatory CD206 − macrophages was higher in the LV of males compared to females, whereas the number of anti-inflammatory (pro-fibrotic) CD206 + macrophages was higher in females vs. male mice. By aging, the collagen I/III ratio in the LV was higher in female mice compared to male mice, which was associated with a higher collagen content in 12-months old female compared to male cFB. The Figure was created with BioRender.com .

Article Snippet: Acetone-fixed and -permeabilized LV transverse OCT sections, blocked with 1% bovine serum albumin in PBS, were stained with mouse anti-α-actinin (A7811, Sigma-Aldrich; secondary antibody: FITC donkey anti-mouse, 715-096-150, JacksonImmunoResearch, West Grove, PA, USA), rat anti-mouse CD68 (MCA 1957GA, Bio-Rad; Hercules, CA, USA; secondary antibody: Alexa Fluor 647 Donkey anti-rat IgG, 712-605-153, JacksonImmunoResearch) and goat anti-mouse CD206 (AF2535, R&D, Wiesbaden, Germany; secondary antibody: Rhodamine Red-X Donkey anti-goat, 705-295-147, JacksonImmunoResearch).

Techniques: Derivative Assay, Migration