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. 2018 Feb 5;215(2):423–440. doi: 10.1084/jem.20171274

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© 2018 Hulsmans et al.

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Figure 2. — Ccr2-dependent monocyte recruitment contributes to cardiac macrophage expansion associated with diastolic dysfunction. (A) Flow cytometric quantification of monocytes and neutrophils in blood from control, SAUNA-exposed, and aged mice. Left: Representative flow cytometry plots; right: number of leukocytes and myeloid cells per milliliter of blood. Data are pooled from 2 (aging) to 11 (SAUNA) independent experiments (n = 8–89 mice per group). (B) Relative expression levels of different chemokines and adhesion molecules by qPCR in hearts from control, SAUNA-exposed, and aged mice. Data are pooled from two (aging) to four (SAUNA) independent experiments (n = 10–33 mice per group). (C) Flow cytometric quantification of myeloid cell populations in hearts from control and C57BL/6 and Ccr2^−/− SAUNA-exposed mice. Left: Representative flow cytometry plots; right: number of cell populations per milligram of heart tissue. Data are pooled from three independent experiments (n = 14–15 mice per group). (D) Immunohistochemical analysis of macrophages in hearts from control and C57BL/6 and Ccr2^−/− SAUNA-exposed mice. Left: Representative images; right: bar graph shows percentage of positive staining per ROI. Data are pooled from two independent experiments (n = 5–12 mice per group). Bar, 25 µm. (E) Relative Anp and Bnp expression levels by qPCR in hearts from control and C57BL/6 and Ccr2^−/− SAUNA-exposed mice. Data are pooled from two independent experiments (n = 8–12 mice per group). (F) Lung wet-to-dry weight ratio in control and C57BL/6 and Ccr2^−/− SAUNA-exposed mice. Data are pooled from two independent experiments (n = 4 mice per group). Results are shown as mean ± SD. For statistical analysis, one-way ANOVA followed by Tukey’s test was performed for multiple comparisons. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.