Ptrka

Protein expression analysis was performed in nodose ganglion samples for calcitonin gene related protein (CGRP) (diluted 1/500) (Biomol, Pennsylvania PA, USA) and in SMG for tyrosine hydroxylase (Th) (dil 1/600), Sema 3A, sortilin (Sort1) (diluted 1/1000) (Abcam, Cambridge, UK), TrkA, Rho kinase (ROCK) (diluted 1/200) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), phospho-TrkA (pTrkA), mitogen activated protein kinase (MAPK), phospho-MAPK (pMAPK), c-Jun N-terminal kinase (JNK), phospho-JNK (pJNK), protein kinase B (AKT), phospho-AKT (pAKT), cleaved caspase7 (casp-7), poly(ADP-ribose) polymerase (PARP) (diluted 1/1000) (Cell Signaling Technology, Beverly, MA, USA) and neural nitric oxide synthase (nNOS) (1/1000) (BD Biosciences, San Jose, CA, USA). See data S1 for details.

The following antibodies were purchased: pTrkA (Tyr⁴⁹⁰; 9141), pALK (Tyr⁶⁶⁴; 3341), pSTAT3 (Tyr⁷⁰⁵; 9131), pIGF‐IR (Tyr¹¹³¹; 3021), IGF‐IR (9750), AKT (9272), and pAKT (Ser⁴⁷³; 4051) (Cell Signaling Technology, Danvers, MA, USA); ALK (M7195; Dako, Carpinteria, CA, USA); ALK (ab17127; Abcam, Cambridge, MA, USA); TrkA (sc‐118), ERK1/2 (sc‐94), caspase‐3 (sc‐7272), BCL‐2 (sc‐7382), and BCL‐X_L (sc‐8392) (Santa Cruz Biotechnology, Santa Cruz, CA, USA); STAT3 (569388; Calbiochem); TrkA (06‐574) and NGF (04‐1119) (EMD Millipore, Billerica, MA, USA); and β‐actin (A‐2228; Sigma).

All chemicals were from Sigma-Aldrich (Saint Louis, MO, USA) unless stated otherwise. Comet assay kits, reagents and slides were purchased from Trevigen (Gaithersburg, MD, USA). Antibodies, including phospho-ERK (pERK), pJNK, pp38, pAkt, pp53, pTrkA, ERK, JNK, p38, p53, ATM, H2AX, Egr-1, NF-M, TrkA, and cleaved caspase-3 were purchased from Cell Signaling Technology (Beverly, MA, USA). The pATM and γH2AX antibodies were obtained from Millipore (Billerica, MA, USA). Akt, LOX-1 and GAPDH antibodies, and siRNA were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Alexa Fluor 488-conjugated antibody and Lipofectamine 2000 reagent were purchased from ThermoFisher Scientific (Waltham, MA, USA). Recombinant human β-NGF and caspase inhibitor Z-VAD-FMK were purchased from R & D System (Minneapolis, MN, USA). Horseradish peroxidase-conjugated anti-mouse and anti-rabbit IgG antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). All cell culture reagents were obtained from Gibco-BRL/Invitrogen (Carlsbad, CA, USA).