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Fig. 10. CRM1 interacts with newly imported snRNPs in vivo. (A) Immunoprecipitations from lysates of cells expressing YFP-SmB (i) or YFP (ii) using anti-GFP antibodies. Detection of products using anti-GFP demonstrates efficient precipitation of YFP-SmB and YFP (GFP Beads lane). Duplicate blots probed with anti-CRM1 demonstrate co-immunoprecipitation of endogenous CRM1 with GFP-SmD1 (iii) or YFP-SmB (iv) after 24 hours of expression (GFP Beads lanes), but not with GFP-SmD1 (v) or YFP-SmB (vi) expressed for 48 hours or YFP alone (vii) (GFP Beads lanes). (B) Immunoprecipitations from lysates of cells expressing CRM1-GFP using anti-GFP antibodies. Detection of products using anti-GFP demonstrates efficient precipitation of CRM1-GFP (i) (GFP Beads lane). Duplicate blot probed with anti-SMN (ii) or Y12 anti-Sm (iii) demonstrates co-immunoprecipitation of endogenous SMN and Sm proteins with GFP-CRM1, whereas the use of anti-U2B'' (iv) demonstrates no co-immunoprecipitation of this marker of mature snRNPs. (C) Immunoprecipitations from cells expressing GFP-SMN or UIA-GFP using anti-GFP antibodies. GFP-SMN (i) co-immunoprecipitates endogenous CRM1, whereas U1A-GFP (ii) does not.

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