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. Author manuscript; available in PMC: 2019 Feb 1.
Published in final edited form as: Mol Cancer Res. 2017 Nov 8;16(2):322–332. doi: 10.1158/1541-7786.MCR-17-0454

Figure 7.

Figure 7

The association of EGFR with TROY enhances TROY-induced NF-κB activation. (A) Schematic representation of the expression construct encoding TROY TRAFm protein. The extracellular domain, transmembrane (TM) domain, cytoplasmic domain, and mutations of TRAF binding sites are indicated and the 3X HA epitope tag is shown. (B) Q293/NF-κB-luc cells were transfected with the indicated plasmids. Twenty-four hours after transfection, cells were serum starved (0.1% FBS) for 16 h and lysed and NF-κB-luc reporter expression was measured using luciferase reporter assay kit. Luciferase activity was normalized to the vector-transfected cells. The values shown are mean ± SD from three independent experiments. ***, p < 0.001. (C) The lysates for the luciferase assay (B) were immunoblotted with the indicated antibodies. (D) Q293/NFκB-luc cells were transfected with the indicated plasmids. Twenty-four hours after transfection, cells were serum starved (0.1% FBS) for 16 h and then left untreated or treated with 20 nM EGF for 90 mins. Cells were lysed and NF-κB-luc reporter expression was measured using luciferase reporter assay kit. Luciferase activity was normalized to the vector-transfected cells. The values shown are mean ± SD from three independent experiments. *, p < 0.05; **, p < 0.01. (E) The lysates for the luciferase assay (D) were immunoblotted with the indicated antibodies.