FIG 2.

The arginine-rich motif (ARM) of E4orf4 is required for interaction with Nup205. (A) FLAG-Nup205 was cotransfected in H1299 cells with wild-type or the indicated point mutants of E4orf4. Immunoprecipitation (IP) from cell extracts was performed using anti-FLAG antibody. IP complexes were analyzed by Western blotting to detect E4orf4 (anti-HA) and Nup205 (anti-FLAG). Anti-HA Western blotting of whole-cell extract (WCE) was used to confirm expression of FLAG-Nup205 (middle portion). (B) H1299 cells were cotransfected with FLAG-Nup205 and empty vector (EV), myc-pyruvate kinase (myc-PK), or myc-PK fused to the E4orf4 ARM (myc-PK-ARM). IP from cell extracts was performed using anti-FLAG. IP complexes were analyzed by Western blotting to detect myc-PK and myc-PK-ARM (anti-myc). myc immunoblotting of WCE is used to confirm expression of PK fusions. The asterisks indicates a background band with anti-myc antibody.