Fig. 8. RXR-TR3 interaction correlates well with their ability to target to the mitochondria. (A) Cellular localization of GFP-TR3N and GFP-TR3C in response to 9-cis-RA. GFP-TR3N or GFP-TR3C together with RXR was transfected into MGC80 cells. Cells were treated with or without 9-cis-RA (1 µM) for 12 hours, then immunostained with anti-Hsp60 antibody. GFP-TR3N, GFP-TR3C and mitochondria (Hsp60) were visualized using a confocal microscope. (B) Interaction of RXR with different TR3 mutants. GFP-TR3N or GFP-TR3C together with Myc-RXR was cotransfected into HEK293 cells. Cell extracts were prepared and immunoprecipitated with anti-Myc antibody. The immunoprecipitate was subjected to SDS-PAGE, blotted and probed with anti-GFP antibody. The same membrane was also blotted with anti-Myc antibody to determine immunoprecipitation (IP) specificity and efficiency. Input represents 5% of cell lysates used in the IP western blot assay. The empty vector was used as a control.