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

Figure 5.

Figure 5.

The minimal ORE of the COX4I2 promoter is maximally active at 4% oxygen. (A) ORE reporter activity at varying oxygen tensions. The 293 cells were transfected with 1 µg of the reporter plasmid and maintained at the indicated oxygen tensions for 48 h, followed by analysis of reporter activity relative to normoxia. (B) Real-time PCR analysis of COX4I2 transcript levels of RBPJ, CXXC5 and CHCHD2 at 4% oxygen. (C) Endogenous protein levels of RBPJ, CXXC5 and CHCHD2 analysed by immunoblotting of whole cell lysates from cells maintained at 20% and 4% oxygen for 48 h. (D) Expression levels of the individual proteins in transfected cells maintained under normoxic and hypoxic conditions for 48 h. GAPDH was used as a loading control (*P <0 .05). (E) CHCHD2 is localized in the nucleus. Nuclear fraction of cells maintained at 20% and 4% oxygen were immunoblotted for the levels of CHCHD2. DRBP76 is the nuclear marker and COX1 is the mitochondrial marker. A representative blot has been shown. (F) Quantitation of the level of nuclear CHCHD2 at normoxia (20% O2) and hypoxia (4% O2).