Fig. 3

Relative gene expression in the lumbar region of the spinal cord. a depicts a diagram of the lumbar enlargement that was taken for RNA extraction, cDNA synthesis, and real-time PCR analysis. Males and females were treated with either water (n = 8 per sex) or minocycline (n = 8 per sex; 50 and 25 mg/kg 12 h later) then treated with morphine (4 mg/kg) or saline s.c. Tissue was collected 20 min post morphine administration. Data were analyzed using a 2 × 2 × 2 ANOVA with sex, morphine treatment, and minocycline treatment as variables. Main effects and interactions are depicted as indicated below. Graphs represent the mean ± SEM. b depicts the analysis of microglial and inflammatory markers of the spinal cord, specifically CD11b, TLR4, and IL-1β. Analysis of CD11b revealed a main effect of sex (^# p < 0.05) and interaction between sex and minocycline treatment (*p < 0.05). c Analysis of the astrocyte marker, GFAP, shows a significant interaction between sex and minocycline treatment (*p < 0.05) and a main effect of sex (^# p = 0.001). d depicts the analysis of the fractalkine receptor CX3CR1 and its ligand CX3CL1. CX3CR1 reveals a main effect of sex (^# p < 0.05)