Figure 9
From: Phospholipase D regulates the size of skeletal muscle cells through the activation of mTOR signaling
PLD regulates mTOR signaling in myotubes. (A) Left panel: myotubes were left untreated (C), or were treated with 0.5 μM FIPI, or with 100 nM of a PLD1-specific inhibitor (i-PLD1), or with 100 nM of a PLD2-specific inhibitor (i-PLD2). Middle panel: myotubes were transfected with control siRNA (siC), or with PLD1-siRNA (siPLD1), or with a siRNA directed against both PLD1 and PLD2 (siPLD1/2). Right panel: myotubes were infected with a GFP-adenovirus as a control, or with PLD1- or PLD2-adenovirus. Protein extracts were used to evaluate Phospho-Thr389/Thr412-S6K1 by immunoblotting. Quantification of the bands normalized to the total S6K1 amount is given in the diagrams, as means ± SE of n = 3 to 5 experiments. (B) Myotubes were treated as described above, and Ph-Ser473 Akt was detected by immunoblotting in the protein extracts. Quantification of the bands normalized to the total Akt amount is given in the diagrams as means ± SE of n = 3 to 7 experiments. **: different from control, p < 0.01; *: p < 0.05.