Figure 3

LFC assay of TrkB homodimers. (A) HEK 293 cells expressing indicated protein(s) were cultured in a microtiter plate, and then incubated with luciferin as described in Methods. The red color of the upper panels shows the intensity of bioluminescence produced by the cells in the microtiter wells, and the lower histogram shows the mean ± S.D. of triplicate experiments. *** p < 0.001. (B) HEK 293 cells expressing a full-length luciferase or co-expressing TrkB-NLuc and TrkB-CLuc were treated with BDNF at indicated final concentrations for 10 min, and then incubated with luciferin. The black color of the upper panels shows the intensity of bioluminescence produced by the cells in the micrtotiter wells. The lower histogram shows the mean intensity ± S.D. of triplicate experiments. (C) HEK 293 cells co-expressing TrkB-NLuc and TrkB-CLuc were stimulated with 100 ng/ml of BDNF in the presence of luciferin and 0.45 M sucrose to prevent endocytosis as described in Methods. Data are the mean ± S.D. of triplicate independent experiments.