Figure 3. Western blots of co-immunoprecipitated wild-type and mutant ELOVL4

The EGFP antibody was used to precipitate wtEGFP/ELOVL4 and mutant ELOVL4 protein complexes. A DsRed monoclonal antibody was used for immunodetection of mtDsRed/ELOVL4 proteins. Lane 1: The EGFP antibody (specific for the wtEGFP/ELOVL4 protein construct) co-immunoprecipitated the mtDsRed/ELOVL4(5bp-del) protein. Lane 2: Untransfected control. Lane 3: Control showing expression of both proteins wtEGFP/ELOVL4 (upper band, about 63 kDa) and mtDsRed/ELOVL4(5bp-del, lower band, about 53 kDa) with co-transfection. IP represents immunoprecipitation; WB represents western blot; anti-EGFP represents antibody to enhanced green fluorescent protein; anti-DsRed represents antibody to red fluorescent protein; wt EGFP/ELOVL4 represents fusion protein containing the enhanced green fluorescent protein fused in frame to the N-terminal end of wildtype ELOVL4 protein; mtDsRed/ELOVL4(5bp-del); fusion protein containing the red fluorescent protein fused in frame at the N-terminal end of a mutant (C-terminal 5bp deletion) ELOVL4 protein.