Fig 2: Developmental regulation of BDNF-induced TrkB receptor phosphorylation.(a) Representative blots of experiments showing age-dependent modification of BDNF-induced (50 ng/ml, 5 min, at 37°C) TrkB tyrosine phosphorylation (Y816) response in mouse cortical (PFC) and hippocampal (HC) microslices ex vivo. (b) Representative blots of experiments showing that BDNF (50 ng/ml, 15 min, 37°C) readily induces TrkB phosphorylation at sites Y515 and Y705/6 in P8 hippocampal microslices whereas neither site is effectively phosphorylated by BDNF in P24 hippocampal microslices. (c) NGF (50 ng/ml, 15 min, at 37°C) readily induces TrkA tyrosine phosphorylation (Y674/5) in P24 hippocampal microslices whereas BDNF (50 ng/ml, 15 min, at 37°C) has no effect on TrkB phosphorylation (Y705/6) under the same conditions. For statistical analysis, a one-way ANOVA followed with Bonferroni post hoc test was performed; ***P<0.001. n = 4 per group. Phospho-TrkB values are normalized against total TrkB levels.

Fig 3: The indicated RET- and TRKA-transformed NIH3T3 cells were inoculated subcutaneously into both flanks of nu/nu mice. After four days, mice were randomly assigned to receive Pz-1 (0.3, 1 or 3 mg/Kg) by oral gavage or left untreated. Average size ± SD of tumours is reported.

Fig 4: Identification of TrkA-1q23.1 genomic amplification in MM patients. a, hotspots of 40 minimum common amplifications (red) in primary MM genome, detected by aCGH across 31 patient samples, are plotted along their corresponding chromosome position and proportionally to the respective amplicon size. Detailed genomic information of hotspots is provided in Additional file 1: Table S2. b, schematic segmental gain profile within the 1q23.1 region (spanning ~2.5 Mb), as defined by aCGH, is represented with horizontal bars, each denoting the copy number status of an individual MM patient. MM samples with increasing primary tumor thickness are at the top (for details see Additional file 1: Table S4). Genomic amplifications are depicted in red. The black boundaries delineate the extent of the minimal common amplification (genomic coordinates chr1:156826196 to chr1:157106439). The graphical layout of the genes localized in the minimal amplification is based on the Ensembl release 75.37 of the human genetic map. The region of minimal common amplification extends over ~280 kb and retains the functional transcript of the NTRK1 gene [GenBank: Y09028], NTRK1-001 (red and inset), which codes for TrkA protein. For each panel, the corresponding scale of genomic positions (in Mb) is indicated. c, box and whiskers graph showing the association of TrkA-1q23.1 minimal amplification and tumor thickness in primary MM samples analyzed by aCGH (n = 31; Mann-Whytney U test: *, p < 0.05). Dipl, diploid copy number; Dup, duplication; Amp, amplification

Fig 5: Proposed binding pose and interactions of Pz-1 in TRKA (A) and RET (B). Kinase structure: yellow ribbons; hinge residues: pink ribbon; DFG residues: blue ribbon; aC-helix: green ribbon; carbon atoms: orange; nitrogen atoms: blue; oxygen atoms: red.