Fig. S1

PI(3)Kγ is necessary for directional migration of neutrophils to wounds(related to Figure 1)

(A) Larvae at 3 dpf were pretreated with LY294002 or DMSO for 1 hour and wounded in the tail fin with a needle. Neutrophils were stained by Sudan Black staining 1h after wounding. The number of neutrophils in the fin (magenta) and at the wound (green) was counted for each condition. Note that LY294002 inhibits recruitment of neutrophils in a dose-dependent manner. n=33 (DMSO), 39 (32.5 μM LY294002), 38 embryos (65 μM LY294002 ), *, P<0.01, two-tailed unpaired t-test.

(B) Larvae at 3dpf were pretreated with AS-605240 or DMSO for 1 hour and wounded in the tail fin with a needle. Neutrophils were stained by Sudan Black staining 1h after wounding. The number of neutrophils in the fin and at the wound was counted for each condition. Inhibition of PI(3)Kγ with AS-605240 significantly inhibits neutrophil attraction to wounds. n=19 (DMSO), 26 (2.5 μM AS-605240), 25 (10 μM AS-605240), *, P<0.01, two-tailed unpaired t-test.

(C) Representative images of fluorescent leukocytes in control or PI(3)K morphants (Tg(MPO:mCherry)uw at 2 dpf). These morpholinos do not affect the location or number of neutrophils.