Effects of AM1241, EA, AM1241 plus AM630, and EA plus AM630 on macrophages labeled with β-endorphin in the skin tissues. A, representative confocal images showing ED1-positive macrophages (ED, green, left panels) and β-endorphin-immunoreactive cells (β-EP, red, middle panels). The right panels are overlay images showing double-labeled β-endorphin and macrophages (Merge, yellow, white arrows). Images include vehicle control (CONT), CFA+vehicle of AM1241 (CFA), CFA+AM1241 (AM1241), CFA+AM1241+AM630 (AM1241+AM630), CFA+sham EA+vehicle of AM630 (sham EA), CFA+EA+vehicle of AM630 (EA), and CFA+EA+AM630 (EA+AM630) groups (Scale bar, 50 μm). B-C, summary graphs show the percentage of macrophages labeled with β-endorphin (β-EP) in the total of ED1-positive macrophages in the skin tissues. Data are expressed as means ± SEM (n = 6 rats in each group). * P < 0.05, compared with the vehicle control group; # P < 0.05, compared with the CFA+vehicle of AM1241 group; & P < 0.05, compared with the CFA+AM1241 group; †P < 0.05, compared with the CFA+sham EA+vehicle of AM630 group; $ P < 0.05, compared with the CFA+EA+vehicle of AM630 group (One-way ANOVA followed by Tukey's test).