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. 2013 Jan 8;41(4):2255–2266. doi: 10.1093/nar/gks1454

Figure 6.

Figure 6.

CXXC5 and CHCHD2 interact with RBPJ in vivo. (A) The 293 cells were transfected with C-terminal His-tag expression plasmids for RBPJ and either CXXC5 or CHCHD2. After 48 h at 20% or 4% oxygen, RBPJ was immunoprecipitated using the anti-His-tag antibody. The eluted protein was immunoblotted and probed using anti-CXXC5 antibody. (B) Immunoprecipitation was performed as described above for cell extracts at 20% and 4% oxygen and the eluted protein was probed for CHCHD2 using the anti-FLAG antibody. The blot was reprobed using the anti-His antibody to show the input fraction. (C) RBPJ interacts with endogenous CXXC5 and CHCHD2. The 293 cells were transfected with an expression plasmid for RBPJ followed by an immunoprecipitation of the nuclear extracts. The elution fraction was probed for the presence of endogenous CXXC5 and CHCHD2. (D) CHCHD2 overrides CXXC5 mediated inhibition under hypoxia. The 293 cells were transfected with 200 ng of the CXXC5, RBPJ and the reporter plasmids along with increasing concentrations of the CHCHD2 expression plasmid. Cells were analysed for reporter activity 48 h after incubation under hypoxic conditions (*P < 0.05).