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. 2008 Aug 27;82(21):10792–10802. doi: 10.1128/JVI.00903-08

FIG. 5.

FIG. 5.

The N-terminal region of CAPERα is required for coactivation with v-Rel. (A) Luciferase assays in cells cotransfected with v-Rel (0.1 μg), CAPERα (1.0 μg) alone or together with excess CAPERα (3.9 μg) or mutant N-CAPERα or CAPERα-C (3.9 μg), along with IL-6κB-luciferase reporter (1.0 μg). The averages and standard deviations from three independent experiments are shown. (B) Coimmunoprecipitation of CAPERα mutants with v-Rel or with wild-type CAPERα. Extracts from 293T cells cotransfected with equal amounts of v-Rel and either CAPERα-Flag, ΔvRID-Flag, or N-CAPERα-Flag DNA (6 μg each) were immunoprecipitated with anti-v-Rel (lanes 3, 6, and 9) or preimmune serum (lanes 2, 5, and 8), followed by immunoblotting with anti-Flag. (C) CAPERα forms homodimers. Coimmunoprecipitation assays were performed in cells cotransfected with equal amounts of Xpress-CAPERα and either CAPERα-Flag, ΔvRID-Flag, N-CAPERα-Flag, or Flag-Mcl-1 control (6 μg each), followed by immunoprecipitation with anti-Flag or IgG and immunoblotting with anti-Xpress. (D) Coimmunoprecipitation assays were performed as for panel C. The blot was reprobed with anti-Flag to determine the amount of immunoprecipitated Flag-tagged proteins. Arrowheads point to immunoprecipitated CAPER-Flag, ΔvRID-Flag, N-CAPERα-Flag, or Flag-Mcl-1 and to low levels of Xpress-CAPER coimmunoprecipitated with N-CAPERα-Flag. Total protein (100 μg) was loaded as input (In).