Atrogin-1/muscle atrophy F-box inhibits calcineurin-dependent cardiac hypertrophy by participating in an SCF ubiquitin ligase complex
J. Clin. Invest. Hui-Hua Li, et al. 114:1058 doi:10.1172/JCI22220 [
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Figure 5Ad-atrogin-1-GFP infection blocks agonist-induced cardiomyocyte hypertrophy. (
A) Cardiomyocytes were infected with Ad-GFP or Ad-atrogin-1-GFP and were stimulated with PE (100 μM) or FBS for 36 hours. Calcineurin phosphatase activity was measured with 5 μg of cell extracts. (
B) Cardiomyocytes were cultured and infected with adenoviruses as described above. The levels of expressed atrogin-1 and endogenous calcineurin A protein were determined by immunoblotting with anti-Myc or –calcineurin A antibodies, respectively. (
C) Cardiomyocytes were cultured and infected with the indicated adenoviruses. Cells were stained with α-actinin antibody (red), and nuclei were stained with DAPI (blue). A representative field is shown for each condition. Magnification, ×200. (
D) Quantitation of cell surface area in
C (100–120 random cells measured in each group).
#P < 0.001 vs. serum-free control; *
P < 0.001 vs. Ad-GFP + PE;
&P < 0.001 vs. Ad-GFP + FBS. (
E) Cardiomyocytes were infected with Ad-siRNA-control or Ad-siRNA-atrogin-1 and were treated with PE for 36 hours. Cells were stained with α-actinin antibody. A representative field is shown for each condition. Magnification, ×200. (
F) Quantitation of cell surface area (100–120 random cells measured in each group).
#P < 0.001 vs. serum-free control; *
P < 0.001 vs. Ad-siRNA-control + PE.
