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. 2007 Sep 10;178(6):913–924. doi: 10.1083/jcb.200702058

Figure 5.

Figure 5.

Analysis of subcellular localization and mobility of wt and mutated GFP-UTF1. (A) A schematic representation and repressor activity of various GFP-UTF1 mutants. The Myb/SANT domain (aa 55–124) and conserved domain 2 (CD2) are indicated by black boxes and gray boxes, respectively. The W63G and E67K point mutations are indicated by asterisks. Repressor activity of the GFP-UTF1 fusion proteins was measured on a constitutively active UAS-TK-Luc reporter in transiently transfected HepG2 cells. Negative controls include pUC18 and a peGFP-C1 plasmid. Error bars represent SD. (B) Confocal images of living cells expressing GFP-UTF1 (1–339), GFP-UTF1 W63G E67K (W63G E67K), GFP-UTF1 1–300 (1–300), and GFP-UTF1 W63G E67K 1–300 (W63G E67K 1–300). (C) FRAP analysis of EC cells expressing either GFP (green line), GFP-UTF1 (1–339; blue line), or GFP-UTF1 W63G E67K (W63G E67K; red line). (D) FRAP experiment of EC cells expressing either GFP (green line), GFP-UTF1 (1–339; blue line), or GFP-UTF1 1–300 (1–300; red line). (E) FRAP experiment of EC cells expressing either GFP (green line), GFP-UTF1 (1–339; blue line), or GFP-UTF1 W63G E67K 1–300 (W63G E67K 1–300; red line). (F) Subnuclear fractionations of stable cell lines expressing GFP-UTF1 (1–339), GFP-UTF1 W63G E67K (W63G E67K), GFP-UTF1 1–300 (1–300), and GFP-UTF1 W63G E67K 1–300 (W63G E67K 1–300). Blots were developed with an antibody against HA. F, free-diffusing protein fraction; D, DNaseI fraction; AS, ammonium sulfate fraction; HS, high salt fraction; M, nuclear matrix fraction. Bar, 15 μM.