Western blot analysis of the PDK1 signaling pathway.
A) Western blot analysis was performed to assess levels of the indicated of proteins and phosphoproteins in PDK1F/F control and PDK1F/F αMHC-Cre knockout mice. B) Expression of total and phosphorylated Akt and Foxo1 was assessed after treatment of cardiomyocytes from neonatal rats with vehicle control (DMSO) or PDK inhibitor (GSK 234470). C) The expression of Nav1.5 in rat cardiomyocytes was assessed after treatment with DMSO, GSK 234470 (GSK), the Foxo1 inhibitor AS 1842856 (AS) or a combination of the two drugs. D-E) Nuclear Foxo1 expression was assessed for cardiomyocytes isolated from control and PDK deletion mice (D) or neonatal rat cardiomyocytes before and after treatment with PDK inhibitor (E). β-actin was tested as a loading control for total cellular protein levels, and histone H3 was tested as a loading control for nuclear protein levels. The mean +SD values from each group were normalized to 1.0 in the control mice. All drugs were dissolved in DMSO and added into cell culture media for 48h before the experiments. In each group, three bands of proteins represented three different hearts. *P <0.05.
