Behavioral responses induced by KGNOP1 and morphine after chronic s.c. administration to mice. (A) Spontaneous locomotor activity in mice treated with saline (control), KGNOP1, or morphine as distance traveled during 30 min after drug daily administration for 4 days. (B) Rewarding effects in mice treated with saline (control), KGNOP1, or morphine in the CPP test after drug conditioning for 4 days. (C) Physical dependence as naloxone-induced withdrawal syndrome in mice treated daily for 5 days with saline (control), KGNOP1, or morphine. Withdrawal was precipitated two hours after last drug administration using naloxone (1 mg/kg, s.c.) and signs of withdrawal were measured over 15 min immediately after naloxone, and a global withdrawal score was calculated. See also Figure S1. Data are presented as the mean ± SEM (n = 8 mice per group). ** p < 0.05, ** p < 0.01, and *** p < 0.001 vs. saline group, two-way ANOVA followed by Bonferroni’s post hoc test (A); * p < 0.05 and ** p < 0.01 vs. saline group, one-way ANOVA followed by Dunnett’s post hoc test (B,C).