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. Author manuscript; available in PMC: 2018 Nov 1.
Published in final edited form as: J Immunol. 2017 Sep 20;199(9):3249–3260. doi: 10.4049/jimmunol.1700823

Figure 6. p205 drives expression from an Asc promoter-luciferase reporter construct.

Figure 6

HEK 293T cells were transfected with a mixture containing 10 ng of TK-Renilla luciferase with 1 ng of Asc promoter firefly luciferase reporter and increasing amounts of (A) p205-HA, p204-HA or Aim2-FLAG, or (B) with full length p205 or p205 deletion mutants as indicated. (C) Transfection of increasing concentrations of c/EBPβ, p65/RelA and p205 alone or, co-transfection of increasing concentrations of p205-HA with either c/EBPβ or p65/RelA with Asc promoter-reporter or, transfection of the Asc promoter-reporter with increasing concentrations of p205 with both c/EBPβ and p65/RelA, as indicated. All luciferase values were measured and normalized to Renilla values. Values are displayed as fold change over the Asc reporter construct alone. Data is representative of three independent experiments. (D) Co-immunoprecipitation and immunoblot of overexpressed HA-tagged p205 and c/EBPβ in HEK 293T cells using anti-HA and anti-c/EBPβ antibodies. (E) Co-immunoprecipitation and Western blot of endogenous p205 and c/EBPβ in LPS-stimulated BMDM using antibodies against p205 and c/EBPβ. Data is representative of two independent experiments (n.s.- non-specific band).