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. 2016 Oct 20;11(10):e0165005. doi: 10.1371/journal.pone.0165005

Fig 3. EZH2 is a transcriptional activator of RelB.

Fig 3

(A) Immunoblots of indicated proteins in SUM-149, MDA-MB-231 or BT-549 cells expressing control siRNA or siRNA targeting p65, RelB or EZH2. β-actin serves as a loading control. (B) Real time PCR showing expression of RelB in SUM-149, MDA-MB-231 and BT-549 cells. Expression was normalized to glucuronidase beta. Values given as means ± standard deviation from three biological repeats. (C) Dual luciferase assay from HEK293 cells co-expressing the RelB Reporter with either Flag-p65 or HA-EZH2 and Renilla luciferase. Relative light units (RLU) was normalized to Renilla luciferase light units and expressed as fold over the RelB promoter alone. Values given as means ± standard deviation of four biological repeats. *P = 0.05, **P< 0.05 versus controls; two tailed unpaired t-test. (D) ChIP analysis of p65 and EZH2 binding at the RelB promoter in SUM-149 and MDA-MB-231 cells. Non-specific binding was estimated from IgG immunoprecipitates. Values given as means ± standard deviation from three technical repeats of one representative experiment from the three biological repeats.