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Figure 3 The effects of nicotine (NIC), 6-hydroxy-L-nicotine (6HLN), and cotinine (COT) (1 and 2 mg/L) administration in scopolamine (SCOP)-treated zebrafish on anxiety-like behavior and locomotor activity evaluated within the novel tank diving test (NTT). (A) The zebrafish locomotion tracking pattern in the NTT according to the experimental groups they belong to. The blue dot represents the beginning of the route while the red dot represents the end of the route. The distance travelled in the top (B) and the time spent in the top/bottom (C) were endpoints for measuring the anxiety, whereas the total distance travelled (D) and the velocity (E) were endpoints for measuring the locomotor activity. The values are expressed as means ± S.E.M. (n = 10). ANOVA analysis identified overall significant differences between the experimental groups for (B) F(8,81) = 21.35, p < 0.0001, (C) F(8,162) = 42.29, p < 0.0001, (D) F(8,81) = 8.555, p < 0.0001 and (E) F(8,81) = 6.172, p < 0.0001. For Tukey posthoc analyses—**** p < 0.0001, *** p < 0.001, ** p < 0.01 and * p < 0.05.