Multiple nucleoporins interact with the N-terminal domain of MX2 (N-MX2).

(A) A yeast two-hybrid (Y-2-H) screen was performed using a human leukocyte cDNA library to identify interacting proteins with wild-type or mutant RRR11-13A N-MX2. Interacting partners were assigned a predicted biological score from A-F to assess the confidence of an interaction being specific (with A indicating very high confidence, and F indicating experimentally determined artifacts). (B) Co-immunoprecipitation of candidate interacting proteins with MX2. 293T cells were co-transfected with HA-tagged wild-type or mutant RRR11-13A MX2 and FLAG-tagged candidate interactors from the Y-2-H screen. Cells were lysed and HA-tagged protein immunoprecipitated with anti-HA antibody. Co-transfection of FLAG-tagged candidates with HA-tagged MX1 or GFP served as negative controls. Immunoblots of immunoprecipitated proteins (IP) were probed with anti-FLAG and anti-HA antibodies. As a control for protein expression, samples of lysate prior to immunoprecipitation (IN) were probed with anti-FLAG antibody. All experiments were performed at least three times.