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After blocking with 5% BSA at room temperature for 1 h, membranes were incubated overnight at 4C with primary antibodies: Akt (Cell Signaling Technology, 9272, 1: 1,000), phosphorylated Akt (Cell Signaling Technology, 9271, 1: 1,000), IGF-1 (Cell Signaling Technology, 73034, 1: 1,000), PI3K (Cell Signaling Technology, 4257, 1: 1,000), phosphorylated PI3K (p-PI3K