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Table S1 Composition of the diets

Table S2 List of antibodies

Table S3 Immunoreactivity in conditioned medium collected from epicardial and subcutaneous adipose tissue explants from guinea pigs

Fig. S1 Effect of conditioned medium from epicardial and subcutaneous adipose tissue on sarcomere shortening in cardiomyocytes. Primary adult rat cardiomyocytes were incubated with control AM or CM (diluted 1:2) from EAT and SAT from SD- and HFD-fed guinea pigs for 30 min. before analysis of contractile function. (A--C). Effect of exposure of cardiomyocytes to CM on departure velocity of contraction (A), peak sarcomere shortening (B) and return velocity of contraction (C). Open bars: AM; grey bars: CM from SD-fed animals; black bars: CM from HFD-fed animals. Data are expressed as mean ± S.E.M. of at least eight independent experiments. Differences between the experimental groups were calculated by one-way ANOVA and unpaired Student's t-tests. #P < 0.05 versus AM; *P < 0.05, HFD versus SD; *P < 0.05; EAT versus SAT.

Fig. S2 Effect of conditioned medium from epicardial and subcutaneous adipose tissue on cytosolic [Ca2+] in cardiomyocytes. Primary adult rat cardiomyocytes were incubated with control AM or CM (diluted 1:2) from EAT and SAT from SD- and HFD-fed guinea pigs for 30 min. before analysis of cytosolic [Ca2+]. (A--C) Effect of exposure of cardiomyocytes to CM on [Ca2+] increase (A), peak [Ca2+] (B) and [Ca2+] decrease (C). Open bars: AM; grey bars: CM from SD-fed animals; black bars: CM from HFD-fed animals. Data are expressed as mean ± S.E.M. of at least eight independent experiments. Differences between the experimental groups were calculated by one-way ANOVA and unpaired Student's t-tests. #P < 0.05 versus AM; *P < 0.05, HFD versus SD; *P < 0.05; EAT versus SAT.

Fig. S3 Contractile function in cardiomyocytes two hrs after removal of CM. Primary adult rat cardiomyocytes were incubated with control AM or CM from EAT and SAT from SD- and HFD-fed guinea pigs for 30 min. Then, the CM were replaced by control AM. Following a 2 hr incubation, contractile function and cytosolic [Ca2+] was analysed. (A--C) Reversibility effect of exposure of cardiomyocytes to CM on departure velocity of contraction (A), peak sarcomere shortening (B) and return velocity of contraction (C). (D--F) Reversibility effect of exposure of cardiomyocytes to CM on [Ca2+] increase (D), peak [Ca2+] (E) and [Ca2+] decrease (F). Black bars: AM; grey bars: cells exposed to CM diluted 1:4; open bars, cells exposed to CM diluted 1:4. Data are mean ± S.E.M. from at least eight independent experiments. Groups were compared by one-way ANOVA and unpaired Student's t-test. #P < 0.05 versus AM; *P < 0.05; HFD versus SD.

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