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. 2014 Dec 8;9(12):e113479. doi: 10.1371/journal.pone.0113479

Figure 5. U12–associated cell cycle distribution in cancer cells.

Figure 5

(A) G1 phase arrest induced by U12 in cell cycle progression of SMMC-7721 cells with various concentrations of U12 for 12 h and 24 h treatment was analyzed using flow cytometric analysis. (B) Western blot analysis of G1 cell cycle regulators (mTOR, p-mTOR Ser2448, p-S6K1 Ser371, p-S6K1 Thr389, PARP, p-Rb Ser807, p-Rb Ser 795, cyclin D1, CDK4, CDK6, and p27) under 24 h of U12 exposure at the indicated concentrations. (C) Western blot analysis of phosphorylated proteins (p- mTOR and p-S6K1 Thr389) within 2 h of 50 µM U12 administration. (D) G1 cell cycle arrest induced by treatment of U12 for 12 h with or without pretreated with rapamycin for 1 h on SMMC-7721 cells. (a, P = 0.479, relative to combination treatment with U12 and rapamycin; b, P = 0.007, relative to combination treatment with U12 and rapamycin). All of the results are representatives from three independent experiments.