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. 2014 Dec 19;111(52):18673–18678. doi: 10.1073/pnas.1413687112

Fig. 1.

Fig. 1.

Generation of tRORγt-TMD, a transducible form of interactomic inhibitor of RORγt. (A) Structure of tRORγt-TMD and its derivatives: nontransducible RORγt-TMD, tRORγt-TMD, a mutant form of tRORγt-TMD without DNA-binding capacity [tRORγt-TMD (RR-AG)], and tRORγt-LBD. (B) Intranuclear transduction efficiency of RORγt-TMD, tRORγt-TMD, tRORγt-TMD (RR-AG), or tRORγt-LBD was examined by Western blot using with anti-FLAG antibody in mouse primary CD4+ T cells after 1-h transduction. (C and D) Dose-dependent (1 h) (C) and time-dependent (D) intranuclear transduction kinetics of tRORγt-TMD was analyzed with nuclear fraction of the cells by Western blot using with anti-FLAG antibody in mouse primary CD4+ T cells. (E) Intranuclear localization of tRORγt-TMD after transduction analyzed by confocal microscopy. (F and G) Functional specificity of tRORγt-TMD was examined in HEK293 cells cotransfected with the vectors expressing wild-type RORγt and luciferase driven by Il17-promoter (F) or with those expressing wild-type RORα1 and luciferase driven by apolipoprotein A5-promoter (G). After 24 h, luciferase activity was analyzed and the value was normalized by Renilla acrivity. Data are representative of at least five (B–D) and three (E–G) independent experiments. Error bars denote SEM. ***P < 0.001.