Fig. 2

Interactions between NS1, cellular proteins and segment 7 mRNAs. a 293T cells were transfected with plasmids encoding GFP or GFP-NXF1 and 48 h later either mock infected or infected with the indicated NS1 mutant viruses at an M.O.I. of 5. Cells were harvested at 6 h p.i. and cell lysates examined by western blotting for the indicated proteins before (total) or after (bound) being subjected to GFP-Trap pulldown assays. b 293T cells were transfected with GFP-NXF1 (GFP-NXF1+) or with GFP alone (GFP-NXF1−) and 48 h later either mock infected or infected with the indicated viruses at an M.O.I. of 10, in duplicate sets. At 6 h p.i., cells were harvested and total cellular RNA was extracted from one set while the second set was subjected to GFP-Trap pulldown assays prior to total RNA extraction. RNA was analysed by reverse transcription with radiolabelled primers specific for segment 7 RNAs followed by urea-PAGE and autoradiography. c, d 293T cells were transfected with either GFP alone or with GFP-tagged NS1 mutant protein expressing plasmids as indicated and 48 h later cells were harvested and subjected to GFP-Trap pull down assays. Eluted samples were loaded onto SDS-PAGE gels and c stained with Coomassie Blue dye or d western blotted for GFP or hsp70 before (totals) or after (bound) GFP-trap fractionation. Lanes 1–3 and 8–10 from a are reproduced with permission from Figure 7 of [4]