Research Paper Volume 2, Issue 1 pp 43—62

Figure 9. SirT1 is necessary for mIGF-1-dependent upregulation of anti-oxidant and hypertrophic genes adiponectin, UCP1 and MT-2. (A) Neonatal primary cardiomyocytes from wild type or mIGF-1 Tg mice were treated with sirtinol (100 μM) or EX-527 (1 μM), or treated with 20 ng/ml IGF-1 for 24 h. (B) HL-1 cardiomyocytes were transfected with the indicated plasmids, or treated with 20 ng/ml IGF-1 for 24 h. Untransfected cells were used as control (CTL). (A, B) The expression levels of adiponectin, UCP-1 and MT-2 mRNAs were examined by Real Time-PCR. (C) Neonatal primary cardiomyocytes from wild type or mIGF-1 Tg mice, and HL-1 cardiomyocytes, were transfected with 1 μg of plasmids carrying Firefly luciferase under the control of promoters of adiponectin (Adipo-Luc), UCP1 (UCP1-Luc) and MT-2 (MT-2-Luc) genes, respectively, together with 1 μg of Renilla Luciferase plasmid. Neonatal primary cardiomyocytes were also treated with different inhibitors or IGF-1 as described in (A). Dual luciferase assays were performed in duplicate for each condition. (A-C) Results are means ± SE of 3 independent experiments (^*,**,***p versus untreated cardiomyocytes).