Fig. 3.

Validation of the TrkA/B/C-expressing neuroblastoma cell system. (A) MYCN protein expression across SH-SY5Y, NBLS and NLF, the parental cell lines used in the study, by Western blotting as shown by Maher et al.³³ (B) Confirmation of TrkA/B/C receptor expression in each isogenic cell line by Western blotting. (Par: parental cell line) (C) Confirmation of Trk receptor activation following ligand stimulation (100 ng/ml; 10 min) of TrkA/B/C expressing cells and downstream activation of Akt and ERK by Western blotting. Cells were serum starved (0.1% FBS) for 6 hours prior to ligand stimulation. Ligands for TrkA, TrkB, and TrkC are NGF, BDNF, and NT-3, respectively. GAPDH acted as loading control for Western blotting.