FIGURE 6:
LST-4::GFP and SNX-1::GFP are transiently localized on maturing phagosomes. All GFP reporters are expressed under the control of Pced-1 promotor. (A and B) GFP and DIC images of wild-type gonad arms (A) or embryos (B) expressing indicated reporters. Arrows indicate phagosomes. Scale bars in (A) and (B), 20 and 10 μm, respectively. (C and D) Time-lapse images displaying LST-4(d)::GFP (C) or SNX-1::GFP (D) on maturing phagosomes in wild-type embryos. “0 min” represents the time point when engulfment is just complete. Scale bars, 2 μm. (E) The temporal order and duration of each fluorescent reporter on extending pseudopods and maturing phagosomes. Data represent means obtained from time-lapse recording of multiple C1, C2, and C3 cell corpses (n, number of phagosomes measured). The light and dark green colors reflect weak and strong signal intensities, respectively. The average phagosomal durations of LST-4(d) and SNX-1 are indicated as mean ± SD. “0 min” represents the time when pseudopod extension is just initiated. (F) Percentage of phagosomes labeled with indicated GFP reporters in 1.5-fold stage wild-type or vps-34(h510); F39B1.1(tm3171) mutant embryos. At least 15 animals were scored for each sample. *, P < 0.001 by independent Student’s t-test. (G and H) Time-lapse images showing the localization of SNX-1::GFP (G) or LST-4(d)::GFP (H) on the tubules extended from phagosomes. Arrowheads indicate GFP-labeled phagosomal tubules. Scale bar: 2 μm.