Figure 2 | Oncogenesis

Figure 2

From: The phosphorylated prodrug FTY720 is a histone deacetylase inhibitor that reactivates ERα expression and enhances hormonal therapy for breast cancer

Figure 2

Nuclear FTY720-P enhances specific histone acetylations in breast cancer cells. MCF7 cells (a) and 4T1 cells (b) were treated with FTY720 (5 μM) for the indicated times. Histone acetylations in nuclear extracts were detected by immunoblotting with antibodies to specific histone acetylation sites. (c) Purified nuclei from naive MCF7 cells were incubated for the indicated times with vehicle, S1P (1 μM) or FTY720-P (1 μM) and histone acetylations determined. (d, e) Purified nuclei were isolated from MCF7 cells transfected with vector, SphK2 or ciSphK2 and treated with FTY720 (1 μM) for 15 min. (f, g) Purified nuclei were isolated from MCF7 cells transfected with siControl or siSphK2 and incubated with the indicated concentrations of FTY720 for 15 min. Histone acetylations were determined by immunoblotting (d, f) and levels of FTY720-P by liquid chromatography, electrospray ionization/tandem mass spectrometry (LC-ESI-MS/MS) (e, g). *P<0.05. (h, i) Naive MCF7 cells were treated with vehicle, FTY720-P (100 nM), FTY720 (1 μM) or SAHA (2 μM) for 2 h, nuclear extracts were analyzed by western blotting with the indicated antibodies (h) and HDAC activity measured and expressed as arbitrary fluorescence units (AFU) (i). *P<0.001.

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