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. 2003 Mar;14(3):836–847. doi: 10.1091/mbc.E02-08-0520

Figure 3.

Figure 3

Sac3 associates physically with mRNA export factors and nuclear pore associated proteins. (A) Sac3 coimmunoprecipitates with Mtr2. Lysates (left) and α-GFP immunoprecipitates (right) from Sac3-myc (PSY2451, lanes 1, 4, 7, and 10), EYFP-Mtr2 (PSY2729, lanes 2, 5, 8 and 11), and double tagged Sac3-myc EYFPMtr2 (PSY2729, lanes 2, 5, 8 and 11), and double tagged Sac3-myc EYFP-Mtr2 (PSY2747, lanes 3, 6, 9 and 12) strains. The asterisk denotes the heavy chain of the α-GFP antibody. Arrows point to EYFP-Mtr2. Lanes 7–9 were exposed ten times longer than lanes 10–12. B. Sac3 coimmunoprecipitates with Mex67. Lysates (left) and α-GFP immunoprecipitates (right) from Sac3-myc (PSY2451, lanes 1, 4, 7 and 10), Mex67-GFP (lanes 2, 5, 8 and 11), and double tagged Sac3-myc Mex67-GFP (PSY2691, lanes 3, 6, 9 and 12) strains. C. Sac3 coimmunoprecipitates with Mlp1. Lysates (left) and α-GFP immunoprecipitates (right) from Sac3-myc (PSY2451, lanes 1, 4, 7, and 10), Mlp1-EYFP (PSY2751, lanes 2, 5, 8, and 11), and double-tagged Sac3-myc Mlp1-EYFP (PSY2752, lanes 3, 6, 9, and 12) strains. (D) Sac3 coimmunoprecipitates with Nup116. Lysates (left) and α-GFP immunoprecipitates (right) from Sac3-myc (PSY2451, lanes 1, 4, 7, and 10), Nup116-EYFP (PSY1832, lanes 2, 5, 8, and 11), and double-tagged Sac3-myc Nup116-EYFP (PSY2856, lanes 3, 6, 9, and 12) strains. (E) Sac3 coimmunoprecipitates with Dbp5. Lysates (left) and α-GFP immunoprecipitates (right) from Sac3-myc (PSY2451, lanes 1, 4, 7, and 10), ECFP-Dbp5 (PSY2726, lanes 2, 5, 8, and 11), and double-tagged Sac3-myc ECFP-Dbp5 (PSY2755, lanes 3, 6, 9, and 12) strains. (F) Sac3 does not coimmunoprecipitate with NLS-NES-GFP. Lysates (left) and α-GFP immunoprecipitates (right) from a Sac3-myc strain containing empty vector (PSY2451 transformed with pPS703; lanes 1, 4, 7, and 10), a wild-type strain expressing NLS-NES-GFP (FY23 transformed with pPS1372; lanes 2, 5, 8, and 11), and a Sac3-myc strain expressing NLS-NES-GFP (PSY2451 transformed with pPS1372; lanes 3, 6, 9, and 12). The asterisk denotes the heavy chain of the α-GFP antibody. Arrows point to two bands corresponding to NLS-NES-GFP. Samples were run on an SDS-PAGE gel. Lysate (10 μg) and 1/20th of the IP (from a total of 1 mg of lysate) were blotted with α-GFP (lanes 7–12) and 10 μg of lysate and the remainder of the IP were blotted with α-myc (9E10, lanes 1–6).