Fig. 2

Dok6 was expressed in DRG neurons. a Dok6 RNA in situ hybridization with tissue from DRG tissues collected at E16.5, E18.5, P3, P7, and P14. Dok6 expression started in the DRG at E16.5, reached a peak at E18.5, and then gradually decreased after birth. The red arrow points to the DRG and the green arrow points to the spinal cord. A Dok6-sense riboprobe was used as the negative control. b Western blot assays for DOK6 with protein lysates from cultured DRG neurons and SCs. DOK6 was predominantly detected in DRG neurons. c qRT–PCR assays with isolated DRG neurons and SCs. Dok6 was mainly detected in DRG neurons. qRT–PCR for Tubb3 and S100β showed a high purity of isolated cells. d Histochemical assays with the DRG neuron-SC coculture system. Cultured cells were subjected to fluorescent in situ hybridization for Dok6 (green), followed by immunostaining for Tuj1 (red) or S100β (red). e Immunostaining of customized antibody to Dok6 in P7 age-matched wild-type control group and mutant mice sciatic nerve and DRG tissue to confirm its expression, Scale bars, 100 μm. f Histochemical assays of the DRG tissue, co-labeling of DOK6(green) with neuronal cell marker Tuj1 (red) and Schwann cell marker S100β (red), Scale bar, 100 μm. g Histochemical assays of the DRG neuron and SC with Dok6. Cultured cells were subjected to immunofluorescence for DOK6 (red), followed by immunostaining for Tuj1 (green) or S100β (green). Scale bar: up panel = 100 μm, low panel: x = 1 μm, y = 1 μm, z = 5 μm. Data are presented as the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, two-tailed unpaired Student’s t test