Fig. 3

Reduction in VCR-induced allodynia by RS-102985 does not require neuronal activation. a Representative images of GFP (macrophages, CX₃CR₁, green) and p-ERK (neuronal activation, red) in L4 DRG taken from VCR-treated CX₃CR₁^+/GFP and CX₃CR₁^GFP/GFP mice at treatment day 11. Mice were co-treated with RS-102985 or vehicle. Scale bar, 50 μm. b Quantification of p-ERK+ profiles per mm² at day 11 (mean ± SEM, n = 4 mice per group). VCR induces neuronal activation, which is significantly reduced by RS-102895 at day 11. ***p < 0.001, one-way ANOVA, Tukey’s test. c Representative blot of p-ERK2 (42 kDa) and α-tubulin (50 kDa) in L3–L5 DRG homogenates obtained from CX₃CR₁^+/GFP and CX₃CR₁^GFP/GFP mice at treatment day 11. d Quantification of p-ERK2 band density normalised to α-tubulin (mean ± SEM, n = 3). In both genotypes, RS-102895 administration significantly reduces VCR-induced p-ERK2 expression in L3–L5 DRG. *p < 0.05, one-way ANOVA, Tukey’s test. e Withdrawal thresholds at days 14–16 remain significantly higher in VCR/RS-102895-treated CX3CR₁^GFP/GFP mice relative to those treated with VCR and vehicle (mean ± SEM, n = 6–9 mice per group). *p < 0.05, two-way RM ANOVA, Tukey’s test. f Representative images of GFP (macrophages) and p-ERK (red) in L4 DRG taken from CX₃CR₁^+/GFP and CX₃CR₁^GFP/GFP mice at day 14. Prior RS-102895 treatment does not continue to prevent p-ERK activation in either genotype. Scale bar = 50 μm. g Quantification of p-ERK+ profiles per mm² at day 14 (mean ± SEM, n = 4 mice per group). NS not significant. h Representative blot and quantification of p-ERK2 and α-tubulin in bilateral L3–L5 DRG homogenates. In both genotypes, DRG from VCR/RS-102895-treated mice express similar normalised p-ERK2 expression to VCR/vehicle-treated mice (mean ± SEM, n = 3)