Figure 1.
Purification of PGC-1α-specific protein complexes, and coimmunoprecipition in the nucleus. HeLa nuclear extract was incubated with purified PGC-1α protein in high salt. Following gradual dialysis and clarification, the complexes underwent sequential chromatography and native elution over Flag and HA affinity columns, respectively. (A) Silver staining showing aliquots of eluted complexes for negative control (no addition of purified protein) and PGC-1α samples following Flag chromatography (left two lanes) and HA chromatography (right two lanes). The arrows designate the positions of p300/CBP, LRP130, and PGC-1α.(B) Subcellular fractionation in H2.35 cells showing colocalization of LRP130 and PGC-1α to the nucleoplasmic fraction. Glycogen synthase (GS) is a cytoplasmic marker. C-V-α is a mitochondrial protein encoded in the nucleus. ND6 is a mitochondrial protein encoded in the mitochondrion. (C) Coimmunoprecipitation demonstrating interaction between LRP130 and PGC-1α in nucleus.