Functional association of Sun1 with nuclear pore complexes -- Data Supplement - JCB--Liu et al. -- The Journal of Cell Biology

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Figure S3. NPC association of various Sun1 isoforms and chimeras. (A) Wild-type p16 MEFs were transfected with HA-Sun1Δ221-343 or HA-Sun1(S3TM) and coimmunolabeled with antibodies against HA and Nup153. High magnification images of the nuclear surface reveal an obvious colocalization of both exogenous Sun proteins with NPCs. (B) To further investigate which domain of Sun1 is involved in NPC targeting, the lumenal domains of Sun1 and 2 were swapped. Sun2(S1L)-GFP contained the nucleoplasmic and TM domains of Sun2 and the lumenal domain of Sun1. Sun1(S2L)-GFP contained the Sun1 nucleoplasmic and TM domains and the lumenal domain of Sun2. Neither chimeric protein localized preferentially to NPCs upon transient transfection in HeLa cells. These data suggest that it is the combination of a portion of the nucleoplasmic domain (residues 1-220) along with the lumenal domain of Sun1 that is responsible for NPC association. Boxed areas are magnified in the panels to the right. Bars, 5 µm.