Figure 5. Colony formation in cancer or normal cells with or without exposure to X-radiation, cell cycle analysis, and AMPK activation in Met- and Mito-Met₁₀-treated PDAC cells.

(A) Cells were treated with Met (left) or Mito-Met₁₀ (middle and right) 24 h before irradiation and clonogenic survival fraction was determined. *, P < 0.05 compared to radiation alone. (B) PANC-1 cells were treated with the compounds for 48 h and cell cycle distribution is shown as a histogram (left) and the percentage of cells in G1 phase (middle). Right panel shows the cyclin D1 protein level in treated cells.*, P < 0.05 and **, P < 0.01. (C, left) Immunoblot of FOXM1 in MiaPaCa-2 cells untreated (vehicle) or treated for 24 h with 1 and 10 μM Mito-Met₁₀ or 1 and 10 mM Met. *, P < 0.05. (Middle, Right) Immunoblots of phosphorylated and total AMPK in MiaPaCa-2 (Middle) and FC-1242 (Right) cells treated with Mito-Met₁₀ and Met. (D) The effect of AMPK inhibition on antiproliferative activity of Mito-Met₁₀. (Left) MiaPaCa-2 cells were pretreated with 0.3 μM Compound C for 48 h, then cell proliferation was monitored in the presence or absence of 0.5 μM Mito-Met₁₀. (Middle) MiaPaCa-2 cells were pretreated with 0.3 μM Compound C for 24 h, then treated in combination with 1 μM Mito-Met₁₀ for another 24 h, and the colonies formed were counted after additional incubation in presence/absence of Compound C as indicated. (Right) The effect of Compound C on the survival fraction. *, P < 0.01 Mito-Met₁₀ alone treatment vs. control. #, P < 0.01 Mito-Met₁₀ alone vs. combination.