, E: quantification of OPN mRNA in WT mice, NHE1 transgenic mice, OPN knockout NHE1 transgenic mice (NHE1·OPN ?/? ), and OPN knockout mice (OPN ?/? ) normalized to ?-actin (n = 10 for WT, n = 9 for NHE1, IL-6) (n = 10 for WT and n = 9 for NHE1)

, quantification of mRNA levels of serum and glucocorticoid-regulated kinase 1 (SGK1) (n = 7 for WT and n = 5 for NHE1). G, H, I: quantification of protein expression in WT and NHE1 mice was performed by Western blot. Specific protein expression was normalized to ?-tubulin

. G. Wt, Bottom left: representative immunohistochemistry images of connexin 43 protein expression in WT and NHE1. Right: quantification of Western blot bands of connexin 43 normalized to ?-tubulin (n = 4 for WT and n = 4 for NHE1). H, left: representative Western blot images of nuclear factor of activated T cells (NFAT) protein expression in WT and NHE1. Immunoblotting was against NFAT (160 kDa) and ?-tubulin (50 kDa), representative Western blot images of connexin 43 protein expression in WT and NHE1. Immunoblotting was against connexin 43 (43 kDa) and ?-tubulin (50 kDa)

?. Fig, A: representative immunohistochemistry images of CD44 expression in the heart cross sections of wild-type mice (WT), NHE1 transgenic mice (NHE1), OPN knockout NHE1 transgenic mice (NHE1·OPN ?/? ), and OPN knockout mice (OPN ?/? ). Bars represent 100 µm. B: quantification of mRNA levels of CD 44 normalized to ?actin (n = 10 for WT, n = 9 for NHE1

, Results are expressed as the ratio of CD44 gene over ?-actin ± SE

?. Fig, A: representative Western blot images of phosphorylated p90RSK protein expression in wild-type mice (WT), NHE1 transgenic mice (NHE1), OPN knockout NHE1 transgenic mice (NHE1·OPN ?/? ), and OPN knockout mice (OPN ?/? ), Role of OPN in NHE1 overexpression-induced phosphorylation of p90 ribosomal S6 kinase

?. Fig, Morphology and histology of heart from wild-type mice (WT), NHE1 transgenic mice (NHE1), OPN knockout NHE1 transgenic mice (NHE1·OPN ?/? ), and OPN knockout mice (OPN ?/? ). A: representative whole heart images. B: representative images of Masson's trichrome staining of heart cross sections. C: quantification of mRNA levels of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) normalized to ?-actin (n = 10 for WT, n = 9 for NHE1, vol.5

, Right: graphical representation of internal and external crosssectional heart cell area (n = 7 for WT, n = 4 for NHE1, n = 4 for NHE1·OPN ?/? , n = 4 for OPN ?/? for internal cell area; n = 8 for WT, n = 5 for NHE1, n = 3 for NHE1·OPN ?/? , n = 4 for OPN ?/? for external cell area). Bars represent 50 µm, Results are expressed as the ratio of target gene over ?-actin ± SE. +P < 0.001 vs. WT, #P < 0.01 vs. WT, ¥P < 0.001 vs. NHE1, ?P < 0.001 vs

, Right: graphical representation of quantification of PSR stain on heart cross sections (n = 14 for WT, n = 10 for NHE1, WT, ?P < 0.001 vs. NHE1 OPN ?/? , £P < 0.05 vs. NHE1, ?P < 0.01 vs

?. Fig, Graphical representation of echocardiographic parameters of wild-type mice (WT), NHE1 transgenic mice (NHE1), OPN knockout NHE1 transgenic mice (NHE1·OPN ?/? ), and OPN knockout mice (OPN ?/? ). Results are expressed as raw values of echocardiographic parameters ± SE, p.1

. Opn-?/? and . ?p-&lt;, B: graphical representation of ejection fraction (EF) and fractional shortening (FS) (n = 10 for WT, n = 9 for NHE1, n = 7 for NHE1·OPN ?/? , n = 10 for OPN ?/? ). C: graphical representation of volumes of left ventricle in systole (LVs) and diastole (LVd) (n = 10 for WT, n = 9 for NHE1, 01 vs. NHE1, ?P < 0.001 vs. NHE1 OPN ?/? . A: graphical representation of the lengths of interventricular septum in systole (IVSs), interventricular septum in diastole (IVSd)