Fig. 7: Dynein participates in DOCK4-modulated trafficking of Nav1.7.

a DRG lysates were immunoprecipitated with DOCK4 antibody and immunoblotted with Dynein and DOCK4 antibody as indicated. This experiment was repeated three times. b DRG lysates were immunoprecipitated with Dynein antibody and immunoblotted with DOCK4 and Dynein antibody as indicated. This experiment was repeated three times. c The Co-IP assay reveals the interaction between DOCK4 and Dynein, with the interaction site identified as the DN (1-655 aa) region of DOCK4. This experiment was repeated three times. d The colocalization of DOCK4 and Dynein in DRG neurons of mice. The box represents magnification. Scale bar, 100 μm. e High-resolution images showing the colocalization of DOCK4 and Dynein in DRG neurons of mice. Scale bar, 10 μm. f Proximity ligation assay (PLA) showed positive signals of DOCK4/Dynein interaction in cultured DRG neurons (6 images from two repeats). Scale bar, 50 μm. g The interaction level between Dynein and Nav1.7 was examined by co-IP in Dock4^WT and AvCreERT2; Dock4^CKO mouse DRG lysates. DRG lysates were immunoprecipitated with Dynein antibody and immunoblotted with Nav1.7 and Dynein antibody as indicated. This experiment was repeated three times. t₄ = 17.34, P = 0.000065. h Proximity ligation assay (PLA) shows positive signals of Dynein/Nav1.7 interaction in cultured DRG neurons of Dock4^WT mice, and signals was loss in cultured DRG neurons of AvCreERT2; Dock4^CKO mice (7 images from two repeats). Scale bar, 50 μm. i Immunofluorescence triple staining demonstrates the co-localization of Dynein, DOCK4, and Nav1.7 in DRG neurons of mice. Scale bar, 100 μm. j High-resolution images show the colocalization of Dynein, DOCK4, and Nav1.7 in DRG neurons of mice. Scale bar, 2 μm. k Changes in membrane expression of Nav1.7 following Dynein knockdown in the DRGs. n = 3 samples per group. t₄ = 7.381, P = 0.0018. l Changes in membrane expression of Nav1.7 following Dynein overexpression in the DRGs. n = 3 samples per group. t₄ = 6.923, P = 0.0023. m–p The effects of Dynein knockdown and overexpression on mouse Hargreaves and tail-flick behaviors. n = 6 mice per group in m and o, n = 5 mice per group in (n and p). t₁₀ = 2.83, P = 0.0179 in m; t₈ = 3.079, P = 0.0151 in n; t₁₀ = 3.054, P = 0.0122 in o; t₈ = 4.236, P = 0.0029 in (p). q Dynein knockdown reversed the effect of DOCK4 overexpression on membrane expression of Nav1.7 in the DRGs. n = 3 samples per group. F_{(2, 6)} = 18.28, P = 0.0034 in control vs. DOCK4-over, P = 0.0071 in DOCK4-over vs. DOCK4-over + Dynein-shRNA. r Dynein knockdown reversed the effect of DOCK4 overexpression on mouse Hargreaves behavior. n = 6 mice per group. F_{(2, 15)} = 6.149, P = 0.0203 in control vs. DOCK4-over, P = 0.0226 in DOCK4-over vs. DOCK4-over + Dynein-shRNA. g, k–p, Two-tailed Independent Student’s t test; q, r, One-way ANOVA followed by Tukey’s multiple comparisons test. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. Data are presented as mean ± SEM. Complete sample size and sex is provided in the Supplementary Data. Source data are provided as a Source Data file.