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. 2010 Nov 15;24(22):2543–2555. doi: 10.1101/gad.1967810

Figure 1.

Figure 1.

RNA-binding protein p68 interacts with insulator-binding protein CTCF. (A) NEs from HeLa cells stably expressing Flag or Flag-tagged human CTCF were used in Flag co-IP experiments and analyzed by immunoblotting with anti-Flag and anti-p68 antibodies. (B) HeLa NEs were precipitated with rabbit IgG or rabbit anti-p68 antibody, shown on the top of the panel, and precipitates were immunoblotted with antibodies, shown on the right of the panel. (C) NIH3T3 NEs were precipitated with mouse IgG or mouse anti-CTCF antibody, shown on the top of panel, and precipitates were immunoblotted with antibodies, shown at the right of panel. (D) Western blot of input, GST-associated, and GST-CTCF-associated proteins isolated from HeLa NEs probed with anti-p68 antibody. (E) Western blot of input, GST-associated, and GST-p68-associated proteins isolated from HeLa NEs probed with anti-CTCF antibody. (F) Schematic representation of GST-CTCF and its deletions used in GST pull-down assays. (G, top panel) GST pull-down assay using immobilized GST only or recombinant GST-CTCF N terminus (GST-CTCF-N), zinc fingers (GST-CTCF-ZF), C terminus (GST-CTCF-C), and full-length (GST-CTCF-FL) proteins probed with anti-p68 antibody by Western blot. (Bottom panel) Ponceau S stain showing total protein present on the membrane before Western blot. Lanes 1 and 2 had 5% and 2% of the input protein samples, respectively. (H) Schematic representation of GST-p68 and its deletions used in GST pull-down assays. (I, top panel) GST pull-down assay using immobilized GST only or recombinant GST-p68 N terminus (GST-p68-N), RNA helicase core region (GST-p68-M), C terminus (GST-p68-C), and full-length (GST-p68-FL) proteins probed with anti-CTCF antibody by Western blot. (Bottom panel) Ponceau S stain showing total protein present on the membrane before Western blot.