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. 2012 Dec 3;32(42):5101–5110. doi: 10.1038/onc.2012.523

Figure 2.

Figure 2

Combined AMPK and FASN inhibition blocks PC cells' proliferation in vitro and induces caspase-dependent apoptosis. (a) Effects of cC and C75 inhibitors on LNCaP and C4-2 cell proliferation was assessed by cell counting at the indicated times. (b–e) Genetic inhibition of AMPK in PNT2, LNCaP and C4-2 cells was performed by nucleofaction with siRNA against hAMPKα1/2. Reduced AMPKα1/2 expression is shown by AMPKα1/2 immunoblotting (b) in LNCaP cells 24 h after transfection with control or siRNA against hAMPKα1/2. (c) Proliferation of sicont- or siAMPKα1/2—PNT2, LNCaP or C4-2 cells was measured by cell counting following treatment with control or C75-supplemented media at the indicated times. (d) Effect on cell viability was analyzed by FACS by measuring the percentage of annexin V-FITC+ cells at 16 h following treatment with control or C75-supplemented media at the indicated times. Rescue from apoptosis induction was analyzed following pre-treatment of cells with 20 μM qVD-oph caspase inhibitor. (e) Effect on cell viability was analyzed by FACS by measuring the percentage of annexin V-FITC+ cells at 16 h after indicated siRNA treatments.