This has been performed as previously described13 (link). Antibodies directed against N-cadherin, and Twist-1 were purchased from Abcam (Cambridge, MA); Cyclin D1 (HD11), β-actin (C-11), p70 S6K (H-9), p-p70 S6K (A-6) and 4E-BP1 A (P-1) were purchased from Santa Cruz (Santa Cruz, CA). Cleaved caspase-3 (Asp175), Cleaved caspase-9 (Asp 315), Cleaved-PARP-1 (ASP 214), Snail (C15D3), mTOR (7C10), P-mTOR (Ser2448, D9C2), GAPDH, E-cadherin (24E10), Akt and P-Akt (193H12) were purchased from Cell Signaling (Danvers, MA, USA).
After protein transfer, the memranes were cut to 2 or 3 pieces based on the molecular weight to allow simultaneous use of different antibodies corresponding to proteins with different molecular weights.
After protein transfer, the memranes were cut to 2 or 3 pieces based on the molecular weight to allow simultaneous use of different antibodies corresponding to proteins with different molecular weights.
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