A colony of RAGE KO mice on a C57BL/6 background was maintained by breeding heterozygous mice, as previously described24. Heterozygous mice were generated by back-crossing RAGE KO (homozygous) mice37 with C57BL/6 wild type (WT) mice. All experiments used thoracic (T1–T4) dorsal root ganglia (tDRG) from homozygous (RAGE KO) mice and their C57BL/6 (wild type) littermates. Mice were genotyped using polymerase chain reaction as previously described24. Some WT mice were injected with either the RAGE antagonist FPS-ZM1 (Calbiochem, Sigma) (3 mg/kg/day, i.p.38) or a corresponding volume of saline (daily, i.p.) for 48 h before starting experiments. All in vitro experiments involving neuronal primary cultures were done with neonatal pups (P0–P5).

This work was approved by the University of Saskatchewan Animal Research Ethics Board (Campanucci: protocols 20090082 and 20150051) and adhered to the Canadian Council on Animal Care guidelines for humane animal use.

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