Molecular analysis of flies selected for aggressive behavior

Herman A Dierick & Ralph J Greenspan

Supplementary Figure 1 (pdf 88K)
Arena assay and aspiration hole.

Supplementary Figure 2 (pdf 48K)
Array expression profiles of gene in Table 2.

Supplementary Table 1 (pdf 124K)
Biological categories of significant genes³² (P < 0.002).

Supplementary Video 1 (mov 972K)
Main male fighting elements in population cage. The video (three clips spliced together) was shot in a population cage (see Fig. 1a) focusing on one single territory at a time (a blue food cup 2 cm in diameter). The first clip shows wing threat behavior where males lift their wings up at a 45° angle in a threatening posture (which has also been referred to as "wings erect" behavior). The second clip shows the most common male fighting element, referred to as charging or lunging, where the territorial male charges at any intruder on the territory. Often this ends in a lunge where the attacking male lifts his front legs and drops down on the attacked male, who will typically be running away and receive the blow on his side or back. Occasionally, an intruder will not run away, resist the attacks and reciprocate the lunging attacks of the territory holder. This is shown in the third clip and often occurs in short repeated bouts. The two males will then lunge at each other and often box, wrestle and tussle in an attempt to gain control over the territory. We refer to this last behavior as escalated fighting. Such escalations are rare and occur in less than five percent of all aggressive encounters^{10, 21}. In the Aggr selected lines they can last for up to 10 minutes.

Supplementary Video 2 (mov 652K)
Main male fighting elements in scaled-down territorial assay (Fig 1c). The movie was shot in scaled-down territorial assay with two males, one mated female and small food territory (eppendorf cap filled with fly food). Three clips are spliced together, showing wing threat (slow motion), charging & lunging and escalation (slow motion).

Supplementary Video 3 (mov 764K)
Main male fighting elements in arena-assay. The movie shows one arena in the arena chamber. Three clips are spliced together. The first clip shows wing threat followed by a charge and holding, where the attacker grabs the wings of the other male. This often leads to a "roll over", which is quantified as the intensity parameter in Fig 2d. The second clip shows a fighting sequence. The third clip shows both males in escalation mode, boxing and tussling.

Supplementary Video 4 (mov 340K)
Abdomen dipping. The movie was shot in the population cage focusing a single territory. Male streaks his genital area over the food surface while walking as if to mark the territory. Six separate clips are spliced together.

Supplementary Video 5 (mov 128K)
Aspiration of escalating males. The movie shows two males escalating on a blue food territory in the population cage. Aspirator comes in from the left and gently sucks up both males from the territory.

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