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. 2016 Sep 6;113(38):E5628–E5635. doi: 10.1073/pnas.1525078113

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Fig. 4. — Endothelial FoxM1 is essential for cardiac hypertrophy and pathological switch of Ace/Ace2. (A) Quantitation of FoxM1 mRNA in mouse hearts after 7 d after sham or TAC operation. (B and C) Coimmunostaining of FoxM1 (red) and Pecam (green) in mouse hearts 7 d after sham or TAC operation. Arrows, endothelial cell nuclei; arrowheads, myocardial cell nuclei. (Scale bars, 10 μm.) (D and E) Western blot analysis (D) and quantitation (E) of FoxM1 proteins in cardiac endothelial cells isolated from mouse hearts 7 d after sham or TAC operation. P value: Student's t test. Error bar: SEM. (F) Quantitation of ventricle–body weight ratio of mice treated with DMSO (Ctrl) and thiostrepton (Thio) after 4 wk of sham or TAC operation. (G and H) Trichrome staining of cardiac fibrosis in mice treated with DMSO (Ctrl) and thiostrepton (Thio) after 4 wk sham or TAC operation. Red, cardiomyocytes; blue, fibrosis. Arrow, interstitial space. (Scale bars, 20 μm.) (I) Echocardiographic measurement of FS of the LV after 4 wk (4W) of TAC. Ctrl, DMSO; Thio, thiostrepton. (J and K) Western blot analysis (J) and quantitation (K) of Ace and Ace2 proteins in the heart of DMSO-treated (Ctrl) and thiostrepton-treated (Thio) mice 2 wk (2W) after sham or TAC operation. (L and M) Coimmunostaining of FoxM1 (red) and Pecam (green) in control (Ctrl) and SclCre^ERT;FoxM1^fl/fl mouse hearts 4 wk after TAC operation with tamoxifen treatment. Arrows, endothelial cell nuclei; arrowheads, myocardial cell nuclei. (Scale bars, 10 μm.) (N) Quantitation of ventricle–body weight ratio in control (Ctrl) and SclCre^ERT;FoxM1^fl/fl (Mut) mice 4 wk after sham or TAC operation. P value: Student's t test. Error bar: SEM. (O) Echocardiographic measurement of FS of the left ventricle of control (Ctrl) and SclCre^ERT;FoxM1^fl/fl (Mut) hearts after 4 wk of TAC. P value: Student's t test. Error bar: SEM. (P and Q) Trichrome staining of cardiac fibrosis in control (P) and SclCre^ERT;FoxM1^fl/fl (Q) mice 4 wk after sham or TAC operation. Original magnification: 200×. Red, cardiomyocytes; blue, fibrosis. (R) Representative LV pressure–volume (PV) loops taken after cardiac catheterization of control (Ctrl) and SclCre^ERT; FoxM1^fl/fl (Mut) mice 4 wk after sham or TAC operation. (S) Quantitation of Ace, Ace2, and Ace/Ace2 mRNA in control (Ctrl) and SclCre^ERT;FoxM1^fl/fl (Mut) heart ventricles after sham or TAC operation. P value: Student's t test. Error bar: SEM.