Figure 1.

Study design. (A) Characterization of the time course of nonevoked pain (NEP) and evoked (mechanical hypersenstivity, heat hypersensitivity) pain-related behavior profiles after plantar incision. (B) Assessment of nonevoked pain was performed by comparing weight-bearing (print area) of the paw at the incision site (ipsilateral) with the nonincised paw (contralateral) (cohort 1, green, INC N = 8, Sham N = 8). Scale bar = 1 cm. (C) The level of DRG projecting to the plantar surface of the hind paw was determined in our C57Bl6/J mouse colony (cohort 2, grey, naïve N = 8). B = bone, asterisk = musculus flexor digitorum brevis. (D) Mice (N = 54) were assessed for NEP at baseline (BL, 24 hours prior) and 24 hours after (POD 1) plantar incision (INC) or Sham. After determining NEP, mice were sacrificed, and ipsilateral dorsal root ganglia (iDRG) were isolated. iDRG were used for quantitative proteome profiling (DIA-MS; cohort 3, blue, INC N = 15, Sham N = 15) or orthogonal validation by DigiWEST multiplex Western blots (DigiWEST; cohort 4, red, INC N = 12, and Sham N = 12). Obtained results from DIA-MS and DigiWEST analysis were integrated by bioinformatic activity prediction at the network level. (E) Orthogonal validation of our data by immunohistochemistry in ipsilateral L4 DRG at the cell level (cohort 5, light blue, INC N = 3, Sham N = 3).