Fig 3. Chronic EtOH-line worms show resistance to intoxication in F3 generation when yoked-lineage groups are compared.

A. Sensitivity to acute intoxication as measured by percent area covered per worm. EtOH-line animals were derived from a F0 generation that underwent chronic ethanol treatment. Each dot represents a group of 8–16 worms that was assayed to measure locomotion in untreated baseline followed by EtOH-treated conditions. Worms covered less area when treated with EtOH relative to baseline. No significant differences were observed for main effects of lineage or generation, nor after posthoc analysis of interactions between lineage and generation (Table 1). B. To focus on how EtOH treatment changed locomotion from baseline, we plotted locomotion values of EtOH-treated worms normalized to baseline. To show which control- and EtOH-line trials were conducted together, yoked values are linked with a gray line. A two-sided, paired t-test revealed a significant difference between normalized locomotion for control- and EtOH-line worms in F3 generation, but not F1 or F2 generations (p = 0.006). C. To visualize how intoxication sensitivity may differ across yoked trials, we plotted yoked difference values, equivalent to the slopes of linked gray lines in panel B. C1. Distributions of yoked difference values were not different than zero for F1 and F2, but significantly higher than zero for F3 animals (One sample t-test, p = 0.006). C2. Distributions from panel C1 replotted in ranked order to highlight shift towards positive values for F3 in comparison to F1 and F2 generations. A-C. Box plots represent top and bottom 25–75% and median.