Trpml is required for hemocyte migration, phagocytic processing, and immune response. (A ) Representative images of hemocytes cultured with fluorescently labeled bacteria and LysoTracker probe. Scale bar: 5 µm. (B) Phagocytic processing quantified as the percentage of hemocytes containing bacteria within at least two acidic vesicles after 1 h of culture (n = 3 independent cultures, n ≥ 20 hemocytes/condition). (C) Larvae/adult females were injected with bacteria and eclosion/survival rate was quantified. Values are normalized against sterile media injection (n = 3 independent experiments, n ≥ 20 animals/experiment). (D) Left: Representative image of a 16 h ± 30 min–APF pupa with removed cuticle to expose the notum. Red square indicates the region of analysis. Middle: Notum with fluorescently labeled epithelium and hemocyte nuclei. Scale bar: 150 µm. Right: Representative tracks of randomly migrating hemocytes. (E) Hemocyte migration speed quantification in physiological conditions (n = 3 independent experiments, n ≥ 2 animals/experiment). (F) Hemocyte migration speed (left) and directionality (right) measured during 1 h after damage (n = 3 independent experiments, n ≥ 2 animals/experiment); t test (*, P < 0.05). A.U., arbitrary units. (G) Representative images of wound area 1 h after ablation showing the recruitment of hemocytes in both control and trpml-deficient conditions. Scale bar: 10 µm. The values in B are shown as scatter dot plot, indicating mean ± SD, and the values in C, E, and F are shown as box and whiskers (5%–95%), whereby mean values are indicated as “+.” In B, C, and E, one-way ANOVA statistical analysis was performed with Tukey's multiple comparisons post hoc test. Statistically equivalent values are represented with the same letter (P < 0.05). n.s., not significant. Hml and Cg stand for hemolectin and collagen Gal4 drivers, respectively.