Figure 2. Knockdown of Dbi induces mechanical hypersensitivity in mice.

A–E, siRNA against Dbi (or a non-targeting control siRNA) were intrathecally injected (2 μg/site) and 48 h later the following tests were performed: mechanical sensitivity (von Frey) test (A), cold allodynia (dry ice) test (B), Hargreaves test (C), Brush test (D), Alligator clip test (E). Bars are mean ± S.E.M.; ** indicates significant difference with p<0.001 for groups indicated by the connector line (unpaired t-test). F, Recovery of mechanical hypersensitivity (von Frey test) induced by the intrathecal siRNA knockdown of DBI with the intrathecal injection of recombinant DBI (10 ng/site). ### indicate significant difference from baseline (p<0.001); **, *** indicate significant difference from time-matched saline group (p<0.01, p<0.001; two-way repeated-measures ANOVA with Sidák post-hoc test). G, Schematic timeline for the viral DRG gene delivery and osmotic mini-pump experiment; inset depicts DRG 8 weeks after injection with AAV9-U6-shDBI-CAG-EGFP virions. H, Mechanical sensitivity was monitored with the von Frey test during 42 days after the DRG injection of AAV9-U6-shDBI-CAG-EGFP virions or GFP control virions (1.1 – 1.2×10¹² vg/ml; 2 μl). *** indicate significant difference from time-matched control group (p<0.001; two-way repeated-measures ANOVA with Tukey’s post-hoc test). I, Similar to H but heat sensitivity was tested with the Hargreaves test. J, K, Mechanical (J) and heat (K) sensitivity was monitored after the implantation of osmotic mini-pumps delivering recombinant DBI to the DRG (200 μM, 0.5 μl/h; see Methods) to the mice pre-injected with the AAV9-U6-shDBI-CAG-EGFP virions.