Trpml is important for lysosome polarization and myosin nucleation in polarized hemocytes. Representative images of hemocytes expressing (A) LifeAct-GFP stained with LysoTracker probe, (B) LifeAct-Ruby and sqh-GFP, and (C) LifeAct-Ruby and tubulin-GFP.(A–C) Both control and trpml1 conditions are shown. The upper panels are untreated, whereas the lower panels correspond to polarized hemocytes treated with 15 µg/ml of 20-hydroxyecdysone (+20E). Scale bar: 8 µm. (A’–C’) Graphical distribution in 360° of (A’) lysosomes, (B’) myosin, and (C’) tubulin, where 0° = 360°, corresponds to the intersection with the longest axis at the leading edge in a polarized hemocyte. Data are presented as mean ± SEM. (D–F) Front 90° signal compared with the rear 90°, revealing asymmetric distribution of (D) lysosomes, (E) myosin, and (F) tubulin (n = 3 independent cultures, n ≥ 2 cells/condition). Two-way ANOVA with Sidak's multiple comparisons test (*, P value < 0.05). n.s., not significant. All data points are shown with a line connecting the rear and front signal intensities of each hemocyte. Bars represent mean relative intensity of signal distributes at the cell rear (red) and front (blue). A.U., arbitrary units; Ecd., ecdysone.
