Fig. 6

Knockdown of CB1Rd on GABA^RVM neurons antagonizes the analgesic and negative mood-relieving effects of EA. A The diagram illustrating the stereotaxic injection of AAV-mDlx-cre-EYFP virus or AAV-mDlx-EYFP viruses into the RVM region of CB1^flox/flox mice. B The representative fluorescence image shows the injection site of the virus. Scale bars = 500 μm. C Representative immunoblots of CB1R protein levels in the RVM. D The CB1R expression levels were normalized to the expression level of β-actin, n = 4 mice per groups, *** p < 0.001 vs. CB1^flox/flox control group, two-tailed unpaired t-test. E The schematic diagram of the experiment design for the conditioned place preference (CPP) test. F The representative tracking maps of CFA-induced CB1^flox/flox mice in the CPP before and after the EA conditioning. G The time spent in the EA-conditioned chamber and CPP score of CFA-treated CB1^flox/flox mice. H The representative tracking maps of CFA-treated GABA^CB1KO mice in the CPP before and after the EA conditioning. G The time spent in the EA-conditioned chamber and CPP score of CFA-treated GABA^CB1KO mice. For G and I, *p < 0.05 vs. pre-EA condition, two-tailed paired t-test, n = 8. (J-K) The effect of CFA and EA on the tactile withdrawal thresholds (J) and the thermal withdrawal latency (K) of CB1^flox/flox and GABA^CB1KO mice. L The schematic diagram of the experiment design for the conditioned place preference (CPP) test. M The representative tracking maps of CCI-treated CB1^flox/flox mice in the CPP before and after the EA conditioning. N The time spent in the EA-conditioned chamber and CPP score of CCI-treated CB1^flox/flox mice. O The representative tracking maps of CCI-treated GABA^CB1KO mice in the CPP before and after the EA conditioning. P The time spent in the EA conditioned chamber and CPP score of CCI-treated GABA^CB1KO mice. For G and I, *p < 0.05 vs. pre-EA condition, two-tailed paired t-test, n = 8 mice per group. Q-R The effect of CCI and EA on the tactile withdrawal thresholds (Q) and the thermal withdrawal latency (R) of CB1^flox/flox and GABA^CB1KO mice. For J-K and Q-R, *p < 0.05, **p < 0.01 and ***p < 0.001 vs. CFA- or CCI-treatment within CB1^flox/flox and GABA^CB1KO group, two-way ANOVA followed by Bonferroni’s post hoc tests, n = 8 mice per group. S-T The representative tracking maps of CB1^flox/flox and GABA^CB1KO mice in the open field test. U-X The total distance moved (U), the ratio of time spent in the center (V), the entries into the center (W), and the average speed (X) of CB1^flox/flox and GABA^CB1KO mice in the open field. n = 12 mice per group. All data are shown as mean ± SEM