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Figure S1 Isoprenaline induced ER stress in a dose- and time-dependent manner in cardiomyocytes. ER stress marker expressions after isoprenaline treatment for the indicated time and dose. Data are expressed as mean ± SEM, n = 3, *P < 0.05 versus 0 μM or 0 h.

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Figure S2 Isoprenaline injection did not directly cause ER stress. (A) and (B) Western blots of GRP78 expression in isoprenaline-injected rats (n = 4).

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Figure S3 The PDE5 blocker sildenafil suppressed increased activation of calcineurin and Akt in rat failing hearts. Results shown are mean ± SEM; n = 3, *P < 0.05 versus control. #P < 0.05 versus isoprenaline (Iso).

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Figure S4 PDE5 inhibitor sildenafil did not affect the level of cAMP in isoprenaline- or TAC-induced failing hearts (n = 4). #P < 0.05 versus saline or sham.

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Figure S5 Effect of isoprenaline (Iso) on phospholamban phosphorylation. H9c2 cells were treated with isoprenaline (20 μM) in a time-course, and PLB phosphorylation was assayed (n = 4). Data are expressed as mean ± SEM. *P < 0.05 versus 0 h.

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Figure S6 Effect of PKG inhibition on ER stress. H9c2 cells were treated with a range of concentrations of KT5823, for 24 h, and GRP78 expression was assayed. Data are expressed as mean ± SEM (n = 3). *P < 0.05 versus 0 μM.

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Figure S7 PDE5 inhibition attenuated ER stress by up-regulating the SERCA2 activation-dependent PKG pathway. (A) SERCA activity. H9c2(2–1) cells were incubated with sildenafil (Sil; 10 μM) and KT5823 (2 μM) for 24 h and activities were then measured. Data expressed are mean ± SEM (n = 6). *P < 0.05 versus control. #P < 0.05 versus sildenafil. (B) H9c2(2–1) cells were treated with SERCA2 specific siRNA (100 nM) for 24h. SERCA2 expression was determined by Western blot (n = 3). (C) to (G) H9c2(2–1) cells were treated with SERCA2 specific siRNA (100nM) and sildenafil (10 μM) for 24 h. (C) and (D) ER stress marker expression was measured by Western blot (n = 3). (E) and (F) P-PLB-Ser16 and PLB expression was determined by Western blot (n = 3). (G) SERCA activity. *P < 0.05 versus control. #P < 0.05 versus SERCA2 specific siRNA. Data are expressed as mean ± SEM.

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