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. Author manuscript; available in PMC: 2022 Aug 12.
Published in final edited form as: J Med Chem. 2021 Aug 3;64(15):11148–11168. doi: 10.1021/acs.jmedchem.1c00507

Figure 5 -. Cell activities of 27.

Figure 5 -

(a) PRMT5 adduct formation using 27. Expi293 cells were transiently transfected for 48 hrs with an HA-tagged PRMT5:WDR77 complex before treatment with 27. Cultures were harvested at 6 hours, PRMT5 complex isolated by HA-affinity, and analyzed for modification by LC/MS. (b) PRMT5-RIOK1 NanoBiT assay in permeabilized cells. 293T cells stably expressing SmBiT-PRMT5 and LgBiT-RIOK1 proteins. (c) NanoBit assay performed in intact cells after treatment with 27 or 30. (d) Left panel: WB analysis of total symmetric arginine dimethylation levels in MTAP−/− HCT116 cells, in response to 27 or 30. Right panel: WB analysis of total symmetric arginine dimethylation levels in MTAP−/− HCT116 cells overexpressing the PRMT5 ADA mutant which is unable to bind to the PBM peptide, with and without KO of endogenous PRMT5.