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. Author manuscript; available in PMC: 2016 Apr 20.
Published in final edited form as: Angew Chem Int Ed Engl. 2015 Feb 27;54(17):5166–5170. doi: 10.1002/anie.201412154

Figure 4.

Figure 4

SGC707 binds to PRMT3 in cells and reduces PRMT3-dependent H4R3me2a. A) SGC707 stabilized PRMT3 in both HEK293 and A549 cells with EC50 values of 1.3 μm and 1.6 μm in PRMT3 InCELL Hunter Assays. RLU, relative light units. B) Western blot analysis of H4R3me2a levels. HEK293 cells were co-transfected with FLAG-tagged PRMT3 (WT) or its catalytically dead mutant E335Q (Mut) and treated with different concentrations of SGC707, as indicated. The color panels below endogenous and exogenous H4R3me2a and H4 panels are overlays of the above panels in which the methylated H4R3 signal is in green and total H4 is in red thus both signals generating yellow. C) Quantitation of SGC707 effect on the endogenous H4R3me2a from the top panels in (B). D) Quantitation of SGC707 effect on the exogenous H4R3me2a from the bottom panels in (B). The graphs represent nonlinear fits of H4R3me2a signal intensities normalized to intensities of GFP or H4 for exogenous and endogenous H4, respectively, and subtracted for the baseline signal from mutant PRMT3. The results are mean ±SEM of three replicates.