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. 2023 Jan 18;8(5):479–496. doi: 10.1016/j.jacbts.2022.10.001

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© 2023 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Myeloid-Specific NLRC5 Deficiency Exacerbated Cardiac Remodeling Induced by Pressure Overload

(A) Gross morphologies of hearts from Nlrc5^flox/flox (Nlrc5^fl/fl) and LysM-Cre/Nlrc5^flox/flox mice after 4 weeks of TAC. (B) The ratios of HW/TL in Nlrc5^fl/fl and LysM-Cre/Nlrc5^flox/flox mice after TAC (n = 10 per group). (C) Representative images of transverse area of cardiomyocytes detected by wheat germ agglutinin staining. Scale bars, 30 μm. The cross-sectional areas of cardiomyocytes were analyzed by ImageJ (NIH) (n = 10 per group). Ten fields per section from each sample were analyzed. (D) Relative mRNA expression of cardiac hypertrophy-related markers Anp and Bnp were assessed (n = 10 per group). (E to G) Echocardiography analysis of fractional shorting (E), ejection fraction (F), and LV mass (G) after 4 weeks of TAC operation (n = 10 per group). (H) Representative Masson’s trichrome staining and quantitative analysis of collagen volume of hearts from Nlrc5^fl/fl and LysM-Cre/Nlrc5^flox/flox mice after 4 weeks of TAC operation (n = 10 per group). Scale bars, 50 μm. (I) Relative mRNA expression of cardiac fibrosis markers Fibronectin and Collagen Iα were measured in heart tissues from Nlrc5^fl/fl and LysM-Cre/Nlrc5^flox/flox mice after 4 weeks of TAC operation (n = 10 per group). (J) Immunohistochemistry staining and quantitative analysis of cardiac fibrosis marker fibronectin in Nlrc5^fl/fl and LysM-Cre/Nlrc5^flox/flox mice with TAC operation (n = 10 per group). Scale bars, 50 μm. (K and L) Representative cytograms of F4/80⁺ CD64⁺MerTK⁺ macrophages in heart tissues (K) and quantitative analysis (L) (n = 10 per group). (M and N) Quantitative analysis of CCR2⁺Timd4⁻ macrophages (M) and Ly6G⁺ neutrophils (N) in hearts from Nlrc5^fl/fl and LysM-Cre/Nlrc5^flox/flox mice after 1 week of TAC operation (n = 10 per group). (O and P) Quantitative analysis of Ly6C⁺ monocytes (O) and Ly6G⁺ neutrophils (P) in peripheral blood from Nlrc5^fl/fl and LysM-Cre/Nlrc5^flox/flox mice after 1 week of TAC operation (n = 10 per group). Data are expressed as mean ± SD. ∗P < 0.05, ∗∗P < 0.01. Data were analyzed using 1-way analysis of variance followed by Tukey’s post hoc analysis. Abbreviations as in Figure 2.