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. 2012 May 23;3(5):518–524. doi: 10.18632/oncotarget.500

Figure 3. Inhibition of type TGFβ receptor kinase enhances Imatinib mediated CML cell death.

Figure 3

(A) MYL cells were synchronised with a double thymidine block, then treated with either TGFβ or SB431542 for 12 hours. Cells were fixed in 70% ethanol, incubated with propidium iodide and cell cycle status analysed by flow cytometry. Data is representative of three separate experiments and was analysed using CFlow Plus software. (B) MYL cells were serum starved in 0.5% serum containing media overnight and treated with either a range of Imatinib concentrations between 0-0.1μM or in combination with SB431542, or with TGFβ and SB431542 alone as controls. Cell viability was assessed using MTS assays. (C) MYL cells were serum starved overnight in 0.5% serum containing media before treatment with 0.3μM Imatinib, 10μM SB431542 or a combination of the two for 6 or 12 hours. Lysates were separated through SDS-PAGE, and subsequent Western blots were probed with anti-PARP or anti-actin antibodies. (D) MYL cells were pre-treated +/− 1ng/ml TGFβ followed by combination treatments with 0.3μM Imatinib, 10μM SB431542, 20μM MG132 or combinations of these as indicated, and cells were incubated for an additional 24hours and cellular apoptosis measured by annexin-V and propidium iodide staining followed by flow cytometry analysis. Data are representative of two separate experiments performed in triplicate.