Figure S1    TAC IVS, mm 0 0.6 1.2 0.3 0.9 TAC LVPW, mm 0 0.6 1.2 0.3 0.9 n.s. TAC LVDd, mm 0 2 4 6 TAC LVDs, mm 0 2 4 1 3 TAC LVFS, % 0 20 40.

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Figure S1    TAC IVS, mm TAC LVPW, mm n.s. TAC LVDd, mm TAC LVDs, mm TAC LVFS, % * n.s. * * * * * Supplementary Figures Comparison of echocardiographic data after TAC among wild type, heterozygous knockout, and homozygous knockout mice. Echocardiography was performed after TAC operation in wild type (GCLM  ), heterozygous knockout (GCLM  ) and homozygous (GCLM  ) mice. n = 10 in each experiment. n.s. indicates not statistically significant. *P < 0.05.

t-Smad3 p-Smad3 A D p- / t-Smad   TGF  GSH-EE * *   * †† †† B TGF  GSH-EE   † † † † 7.5 † † C   CTGF mRNA expression Procollagen III mRNA expression Procollagen I mRNA expression   TGF  GSH-EE   TGF  GSH-EE     * †† †† † † * †† †† Smad3 phosphorylation and expression of CTGF and procollagen in cultured mouse cardiac fibroblasts The cultured cardiac fibroblasts were incubated with TGF-  1 (25 ng/mL) or vehicle in the presence or absence of GSH-EE (2 mmol/L). (A), Smad3 phosphorylation after the incubation for 24 hrs. Upper panel shows representative blotting. (B-D), mRNA expression of CTGF and procollagen I and III after the incubation for 24 hrs and 4 hrs, respectively. The mRNA levels were normalized to GAPDH mRNA expression. n = 10 in each experiment. +/+ denotes GCLM +/+ mice and  denotes GCLM  mice. *P < †P < The data were expressed relative to the GCLM +/+ fibroblasts treated with vehicle in the absence of GSH-EE (= 1). Figure S2

A TGF-  R2 mRNA expression Sham TAC +/+  * B Sham TAC +/+  TIMP-2 mRNA expression D F/F /+  Time (second) F * GSH / GSSG * * PBS GSH PBS GSH +/+  Figure S3 Myocardial expression of TGF-  R2 and TIMP-2 and microvessel density after TAC, transients of intracellular calcium concentration in cultured cardiomyocytes and effect of GSH-EE supplementation on myocardial microvessel density and GSH/GSSG ratio. (A, B), TGF-  R2 and TIMP-2 mRNA expression. The expression levels were normalized to GAPDH mRNA expression and are shown relative to the levels in sham-operated GCLM +/+ myocardium (= 1). (C), Microvessel number per mm 2 in myocardium after TAC. (D), Representative tracing of transients of intracellular calcium concentration in cultured cardiomyocytes in response to electrical stimulation. (E, F), Effects of GSH-EE supplementation during TAC on myocardial microvascular density and GSH/GSSG ratio.  /  denotes GCLM  /  mice, and +/+ denotes GCLM +/+ mice. n = 6 in each experiment. *P < * Sham TAC +/+  Vessels / mm 2 x 1000 C * * PBS GSH +/+  No Treat. Vessels / mm 2 x 1000 E

ABC Heart rate LVEDP TAC n.s. ** +/+ with no treatment  with PBS  with GSH-EE bpm mmHg * * EF Max dp/dtMin dp/dt TAC * * mmHg/sec D Tau TAC * * ms Systolic BP at aortic root n.s. Figure S4 Effects of GSH-EE supplementation on hemodynamic data 4 weeks after TAC operation in GCLM  mice. Hemodynamic study was performed in GCLM  mice after treatment with GSH-EE or PBS as a placebo and in GCLM +/+ mice with no treatment for 4 weeks after TAC operation. n = 6 in each experiment. n.s. indicates not statistically significant. *P < 0.05.