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. 2010 Jun 14;30(16):3929–3942. doi: 10.1128/MCB.00087-10

FIG. 6.

FIG. 6.

Interaction of Gfi1 with the p65 subunit of NF-κB in NIH 3T3 transfected cells. (A) Schematic representation of full-length Gfi1 (I) and four Gfi1 mutants (II, III, IV, and V) used for coimmunoprecipitation after transfection of constructs into NIH 3T3 cells. Gfi1 deletion mutants Gfi1 I (positions 1 to 257) and Gfi1 III (positions 49 to 257) lack the C-terminal zinc finger domain (gray box), and Gfi1 mutants III and IV lack the N-terminal SNAG repressor domain (black box). (B) NIH 3T3 cells were transiently transfected with Flag-tagged full-length Gfi1 (I) or the Gfi1 mutants (II and IV) and Myc-tagged p65, as indicated, and input levels were controlled by immunoblotting (IB) with anti-Myc or anti-Flag antibodies. (C) Whole-cell lysates of the transfected cells were subjected to coimmunoprecipitation (IP) with anti-Flag antibodies, followed by immunoblotting (IB) with anti-p65 antibodies. (D) NIH 3T3 cells were transiently transfected with the indicated Flag-tagged Gfi1 mutants (II, III, IV, and V) and Myc-tagged p65, as indicated, and input levels were controlled by immunoblotting (IB) with anti-Myc or anti-Flag antibodies. (E) Whole-cell lysates of transfected cells were subjected to coimmunoprecipitation (IP) with anti-Flag antibodies, followed by immunoblotting (IB) with anti-p65 or anti-Flag antibodies.