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. 2015 Jan 6;465(Pt 2):271–279. doi: 10.1042/BJ20141165

Figure 6. Bosutinib and dasatinib fail to induce IL-10 production in macrophages expressing a drug-resistant mutant of SIK2.

Figure 6

The SIK2[T96Q] mutant is insensitive to (A) bosutinib and (B) dasatinib in vitro. Kinase assays were performed using purified SIK2 wild-type and SIK2[T96Q] in the presence of increasing amounts of inhibitors as described in the Materials and methods section (mean±S.D., n=2). (C) Statistical analysis of IC50 curves generated in (A) and (B). (D) RAW264.7 macrophages engineered to express HA-SIK2-WT and HA-SIK2[T96Q] were treated with doxycycline for 16 h, lysed and immunoblotted with the indicated antibodies. Cells transduced with a retrovirus containing no insert was used as negative control (−). (E and F) RAW264.7 macrophages expressing HA-SIK2-WT and HA-SIK2[T96Q] were treated with bosutinib, dasatinib, HG-9-91-01 or vehicle control for 1 h and then stimulated with LPS for 1 h. mRNA expression of IL-10 (E) and nurr77 (F) were measured by qPCR (mean±S.E.M., n=4). WT, wild-type.