Fig. S4

Photoactivation of PA-Rac (related to Figure 4)

(A) Laser stimulation with 458nm light does not have any obvious effects on morphology or migration of neutrophils expressing only mCherry in Tg(MPO:mCherry)^uw. The circles indicate positions of laser stimulation.

(B) (C) Photoactivation of PA-Rac disturbs attraction of a neutrophil to a wound in the fin of Tg(MPO:mCherry-PA-Rac1)^uw. The green and blue lines are tracking of the neutrophil before and after photoactivation. The circles indicate positions of laser stimulation.

(D) Photoactivation of PA-Rac enables guiding of a neutrophil between a wound and the blood vessel, indicating localized Rac activation can overcome endogenous chemotaxis signaling. The trajectory of the neutrophil draws an “apple”.

(E) Photoactivation of PA-Rac is sufficient to induce directed migration of relatively immotile neutrophils in the CHT. Laser stimulation with 458nm light induces migration of a neutrophil with mCherry-PA-Rac (left), but not with mCherry (right). Repetitive photoactivation was performed 10 minutes after starting the imaging (the circles indicate position of photoactivation). To make the kymograph, Z-stack images were summed into a single two-dimensional image, and then consolidated into a semi-one-dimensional line. Scale bars, 10 μm (A, C, E), 50 μm (B), 20 μm (D).