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. 2012 Sep 18;40(21):10689–10700. doi: 10.1093/nar/gks857

Figure 4.

Figure 4.

Sgf11 is distributed between the nucleus and cytoplasm, co-localizes with NPC and interacts with AMEX. (A) Immunostaining of Drosophila S2 cells with antibodies against Sgf11 or Nonstop (red) and NPC (green) and merged images (magnification, ×1000). Panels c and d show cell fragments at higher magnification (×10000). (B) Sgf11 and Nonstop distribution in S2 cell nuclei and cytoplasmic extracts analyzed by western blotting. Equal amounts of extracts were loaded, and the blot was stained with antibodies against indicated proteins. Antibodies against TBP and β-tubulin were used to confirm that the cytoplasmic fraction was not contaminated with nuclear proteins, and vice versa. (C) Sgf11, but not Nonstop, interacts with Xmas-2, a component of the mRNA export complex AMEX, in co-immunoprecipitation from nuclear extract of Drosophila embryos. Antibodies against Xmas-2, Sgf11, Nonstop and IgG (control) were used. Western blot was stained with antibodies against Xmas-2. (D) Sgf11 RNAi knockdown does not affect the level of AMEX components in S2 cells. The presence of the indicated factors was analyzed in cells treated with control nonspecific dsRNA (GFP) or corresponding dsRNA. Tubulin was used as loading control. The levels of corresponding proteins in sham-treated cells (left lanes) or in cells after RNAi treatment (right lanes) are shown.