Supplementary Figure 1: FTY720 is phosphorylated by nuclear SphK2 and enhances histone acetylations in HeLa cells. | Nature Neuroscience

Supplementary Figure 1: FTY720 is phosphorylated by nuclear SphK2 and enhances histone acetylations in HeLa cells.

From: Active, phosphorylated fingolimod inhibits histone deacetylases and facilitates fear extinction memory

Supplementary Figure 1

(a-c) HeLa cells transfected with vector, SphK2, or catalytically inactive SphK2G212E (ciSphK2) were treated without or with FTY720 (5 μM) for 6 h (N = 3 per group). Nuclear and cytoplasmic levels of FTY720-P (a) and S1P (b) were determined by LC-ESI-MS/MS. Data are means ± s.d. *P < 0.01 compared to vector. #P < 0.01 compared to untreated (unpaired two-tail t test). (c) Histone acetylations in nuclear extracts were detected by immunoblotting with the indicated antibodies. (d) Purified nuclei from HeLa cells were incubated for the indicated times with vehicle, S1P (1 μM), or FTY720-P (1 μM) and histone acetylations determined. (e) Naïve HeLa cells were treated with vehicle, S1P (100 nM), or SAHA (1 μM) for the indicated times and proteins were analyzed by western blotting.

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