Table 3.

Stimulatory effects of wild‐type NGF and NGF mutants on activation of TrkA and the subsequent recruitment of the SHC1 protein in recombinant U2OS‐TrkA/p75‐SHC1 cell line

U2OS‐TrkA/SHC1 cells	Interaction between TrkA and SHC1 EC50 (ng/ml) Mean ± SEM
Wild‐type NGF	4.2 ± 1.0
NGF‐R100E	0.56 ± 0.21
NGF‐W99A	23 ± 7.5
NGF‐K95A/Q96A	20 ± 7.7

Activation of TrkA was analysed by measuring the phosphorylation of TrkA at position Y490 and the concomitant docking of SHC‐1 to TrkA‐pY490 after 3 hr of treatment with wild‐type NGF or NGF mutants in the U2OS‐TrkA/p75‐SHC1 cells using the DiscoverX PathHunter assay. EC₅₀ values (ng/ml) are reported as the mean EC₅₀ ± SEM from five independent biological repeats, each with four technical repeats.