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Fig. 2. Analysis of interactions of GFP-tagged SF3a subunits with endogenous subunits and the 15S U2 snRNP. (A-D) HeLa cells were transiently transfected with plasmids encoding full-length GFP-tagged SF3a subunits (FL) or GFP-SF3a60 and GFP-SF3a66 deleted for their zinc finger domains ({Delta}Zn) as indicated above the figure. SF3a complexes were precipitated from small-scale nuclear extracts prepared 48 hours post-transfection with Protein G-Sepharose-coupled anti-GFP (lanes 2-6). Bound proteins were analysed by western blotting for the presence of SF3a60 (A), SF3a66 (B), SF3a120 (C) and SF3b155 (D). Lane 1 shows the extract prepared from cells overexpressing GFP-3a120-FL as an example for the input of the immunoprecipitation reactions. Lane 7 shows the same extract treated with Protein G-Sepharose in the absence of anti-GFP. Positions of precipitated proteins are indicated on the right. The migration of molecular mass standards (in kDa) is shown on the left.





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