Fig. 2

Indo5 is a potent inhibitor for Trk kinase. (a) SK-N-SH cells were pretreated with the indicated doses of Indo5 for 2 h and then treated with NGF (500 ng/ml) for 5 min. HepG2 cells were pretreated with Indo5 for 2 h and then treated with BDNF (50 ng/ml) for 5 min (b). Total cell lysates were prepared for Western blot analysis with the indicated antibodies. (c) Ba/F3-TrkB cells were pretreated with Indo5 for 2 h and then treated with BDNF (50 ng/ml) for 48 h. Cell proliferation was measured and the proliferation index was calculated. The expression of TrkB in Ba/F3-TrkB cells was confirmed by Western blot (right panel). The inhibitory effect of Indo5 on the BDNF-induced TrkB signaling pathway was investigated (d). (e) HepG2 cells were pretreated with Indo5 for 2 h followed by BDNF (50 ng/ml) treatment for 24 h. The number of cells that migrated was measured 24 h later. Data are shown as the mean ± S.D. and are representative of three independent experiments. ** p < 0.01, *** p < 0.001. (f) A549 cells were pretreated with Indo5 for 2 h and then treated with 100 ng/ml EGF (left) or 100 ng/ml VEGF (right) for 5 min. Cell lysates were prepared for Western blot