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. Author manuscript; available in PMC: 2011 Mar 1.
Published in final edited form as: Cancer Cell. 2010 Sep 14;18(3):268–281. doi: 10.1016/j.ccr.2010.08.006

Figure 2. Hes1 enhances NF-κB-dependent activity by inducing IκBα degradation.

Figure 2

(A) Luciferase assay with the 3xκB-luc-reporter in HEK-293T cells transfected with a doxycycline-inducible Hes1 construct. Induction of Hes1 after doxycycline treatment is shown in the lower panel. (B) Immunofluorescence of HEK-293T transfected with flag-Hes1 showing the expression pattern of transfected Hes1 and endogenous p65. (C) Western blot of endogenous IκBα in HEK-293T cells transfected with inducible Hes1 in the presence or absence of TNFα. (D) Lack of IκBα immunostaining in cells transfected with ectopic Hes1. (E) Quantitative RT-PCR to determine the expression levels of IκBα mRNA in the presence or absence of Hes1. Error bars denote +/− Standard Deviation. (F) Western blot of IκBα levels in HEK-293T transfected with the doxycycline-inducible Hes1 construct in the presence or absence of the proteasome inhibitor MG132. (G) Western blot showing the levels of IκBαwt or IκBαK21-22R cotransfected with inducible Hes1. See also related figure S2.