Figure 3
dTAG-13 continuously induced the ternary complex mainly in the cytoplasm. 293A cells coexpressing Ash-FKBP12F36V and CRBN-AG fusion proteins were prepared. (a) Cells were treated with 0.1% DMSO (upper) or 0.1 µM dTAG-13 (lower) for 1 h. Cells were fixed with formalin containing Hoechst 33342, followed by confocal microscopic analysis. AG (green) and Hoechst 33342 (blue) images were merged. Data shown are representative of three independent experiments. Scale bar indicates 50 µm. (b) Numbered magnified areas (i, ii) highlighted with white boxes in (a) are shown. (c) Cells were treated with dTAG-13 (0.001, 0.01, 0.1 or 1 µM) in 0.1% DMSO for the indicated time. Upper panel shows the time course of foci formation until 24 h. Magnified area of the early period highlighted with a gray dashed box in the upper panel is also shown in the lower panel. The number of foci per cell was quantified as in Fig. 1b. Data shown are the mean (n = 4) ± S.D. and are representative of two independent experiments.
