The Divider Assay is a high-throughput pipeline for aggression analysis in Drosophila.

Nevertheless, we detected strong highly significant correlations between manual scoring and automated quantification for our classifier ( R 2 = 0.98, p < 0.0001, n = 60) as well as CADABRA ( R 2 = 0.87, p < 0.0001, n = 59) (Fig. 1d ) showing that correlation analysis is not sufficient to evaluate assay performance.

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In contrast, CADABRA (red boxplots) showed a trend to overscore very low fighting pairs (Fig. 1c , Kruskal–Wallis ANNOVA with Dunn’s test, 0–20, p = 0.17, n = 14) and significantly underscored moderate to very high fighting pairs (Fig. 1c , Kruskal–Wallis ANNOVA with Dunn’s test; 21–100: p = 0.040; 101–300: p < 0.0001; >300, p < 0.0001, n = 15 per group).

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A fivefold increase in arena surface decreased aggression although the decrease did not reach statistical significance.

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