Functional interaction between co-expressed MAGE-A proteins
Fig 3
MageA6 associates with MageA11 in cells.
(A) Co-immunoprecipitation of MageA11 and Mage-A proteins from LNCaP cells. Specific anti-MageA11 antibody was used to immunoprecipitate MageA11. Western blot was performed using a pan-MAGE-A antibody. Anti-GFP was used as IP control antibody. (B) Co-Immunoprecipitation of Flag-MageA11 and HA-MageA6 form HEK293T cells. Immunoprecipitated protein complexes obtained with the anti-Flag antibody (IP anti-Flag) were assessed by Western blot and probed with the indicated antibodies. TL, total lysates. (C) Similar to A but using Flag-MageA11 and GFP-MHD-MageA6. (D) Upper panel: Reporter gene assay for AR activity in HEK293T cells using the PSA-Luc reporter construct and the plasmid encoding AR. MageA11, MageA6, MageA2 and MHD-MageA6 or MHD-MageA2 were co-transfected as indicated in the figure. Cells were treated with DHT 24 h prior to harvest. ev, empty vector. Bottom panel: A sketch of Mage-A protein domains and their expected MW. Error bars indicate mean S.D. Student’s t test was used for statistical analysis. * p < 0.001; ** Immunoglobulin heavy chain. *** Immunoglobulin light chain. Triangles show the corresponding protein band and dashes mark the MW.