Figure 9: Terpene mechanism confirmed by A_2AR CRISPR knockdown in the spinal cord.

Male and female CD-1 mice had CIPN induced along with A_2AR-targeted CRISPR or Negative Control (NC) CRISPR injections into the spinal cord so that day 8 of CIPN and day 10 of CRISPR coincided (see Methods). Data shown is the mean ± SEM, performed in 2–3 technical replicates for each experiment, with sample sizes noted in each graph. BL = baseline. Terpene injected (200 mg/kg, IP), with the exception of β-pinene due to lack of intrathecal response above, followed by mechanical allodynia measurement. A) Time course data shown. *, **, **** = p < 0.05, 0.01, 0.0001 vs. same time point A_2AR-CRISPR group by RM 2 Way ANOVA with Sidak’s post hoc test. B) AUC data calculated from A and compared. * = p < 0.05 vs. same terpene NC group by Unpaired 1-Tailed t Test. The comparison is justified by the independent nature of each experiment along with the demonstrated reduction in antinociception with A_2AR-CRISPR treatment shown in A.