Figure 6. TIM is primary cargo for IMPα1.

(A) S2 cells were transiently transfected with pIZ-tim-V5 (wt), pIZ-tim_mNLS-V5 (tim carrying a mutant NLS), pIZ-imp_α1-VSV, or pIZ-imp_α1∆IBB-VSV as indicated. After 60 hours, cell lysates were immunoprecipitated with an anti-VSV antibody (against IMPα1 or IMPα1IBB) and detected with an anti-V5 antibody. Similar results were obtained in three independent experiments. (B) S2 cells were transiently transfected with pIZ-tim-V5, pAc-per-HA, pIZ-imp_α1-VSV, or pIZ-imp_α1∆IBB-VSV as indicated. After 60 hours, cell lysates were immunoprecipitated with an anti-HA antibody (against PER) and detected with an anti-V5 antibody or with an anti-VSV. Similar results were obtained in three independent experiments. IMPα1 co-immunoprecipitated with PER is indicated with an asterisk. (C) S2 cells were transfected with pIZ-tim-V5 (wt), pIZ-tim ^PL-V5, or pIZ-tim ^TA-V5 in the presence or absence of pIZ-imp_α1∆IBB-VSV as indicated. After 60 hours, cells were subjected to IP using an anti-VSV antibody (against empty vector or IMPα1∆IBB) and detected with an anti-V5 antibody. Similar results were obtained in three independent experiments. The quantification of interaction between IMPα1 and TIM^WT/PL/TA is shown in S9 Fig.