The magnitude of pain after SNI does not differ between sexes following SNI, however; the mechanisms during its onset at the level of the lumbar (L3–5) DRGs are. Spared-nerve injury was performed on the left hindlimb of male and female mice. A, Hind paw mechanical withdrawal thresholds were measured prior to surgery and on days: 1, 3, 5, and 7 post-surgery in both male and female sham (n=7) and SNI (n=7) wild type mice. B, Data for males and females combined shown as mechanical effect size. C, Hind paw response to application of acetone were measured prior to surgery and on days: 1, 3, 5, and 7 post-surgery in both male and female sham (n=7) and SNI (n=7) mice. D, Data for males and females combined shown as acetone effect size. E, Naïve Nav1.8tdT female and male lumbar (L3–5) DRGs were immunostained with Nav1.8 (green), tdTomato (red) and NF200 (Blue); representative images from n=4 mice). Scale bar: 50 μm. Magnification: 20x. F, Quantification of colocalization between Nav1.8, tdTomato and NF200 (n=4). G-H Female and male WT lumbar DRGs (L3–5) were immunostained 3D post SNI with DAPI (teal), Nav1.8 (green), ATF3 (red) and NF200 (blue; representative images from n=4 mice). Scale bar: 50 μm. Magnification: 20x. K, Quantification of ATF3 colocalization with Nav1.8+ neurons of both sexes 3 days after SNI (n=4 per group). L, Quantification of ATF3 colocalization with NF200+ neurons of both sexes 3 days after SNI (n=4 per group). I-J, Female, and male WT lumbar DRGs (L3–5) were immunostained 3D post SNI with DAPI (teal), Nav1.8 (green), HMGB1 (red) and NF200 (blue; representative images from n=4 mice). White arrows point to an example of nuclear or cytosolic localization of HMGB1. Scale bar: 20 μm. Magnification: 40x. M, Quantification of HMGB1 cytosolic localization in Nav1.8+ neurons of both sexes 3 days after SNI (n=4 per group). N, Quantification of HMGB1 cytosolic localization in NF200+ neurons of both sexes 3 days after SNI (n=4 per group). *p < 0.05; **p < 0.01; ****p < 0.0001. BL = Baseline.