Figure 6.
Figure 6. Loss of gop-1 affects phagosomal recruitment of UNC-108. (A and B) Time-lapse images of somatic cell corpses in wild type (WT; A) and gop-1(tm5384) (B) expressing mCherry::MTM-1(C378S) and GFP::UNC-108. 0 min, time point when MTM-1(C378S) was first detected around the cell corpse. The time period until the cell corpse was surrounded by MTM-1(C378S) was defined as the pseudopod extension stage. Quantification of pseudopod extension and the phagosomal duration of MTM-1 and UNC-108 is shown at bottom (mean ± SD). n, number of phagosomes quantified. Yellow arrowheads, MTM-1 release from phagosomes; white arrowhead, recruitment of GFP::UNC-108 to the phagosomal membrane. (C and D) Time-lapse images of germ cell corpses in wild type (WT; C) and gop-1(tm5384) (D) expressing GFP::UNC-108 and mCherry::H2B (Histone 2B). 0 min, time point when phagosomal degradation of the apoptotic cell, indicated by diffuse H2B (white arrowheads), was observed. In wild type, UNC-108 was enriched on the phagosomal membrane 36 min before diffuse H2B was observed, whereas it was absent from phagosomes in gop-1(tm5384). Percentage of phagosomes with the representative pattern is shown in parentheses. n, number of phagosomes that were followed and quantified. In A–D, pink arrowheads indicate the first time point when the intensity of GFP::UNC-108 on the phagosomal surface (white dashed line) was measured. Bars, 5 µm. (E) Cartoon showing a cross section of a pair of gonadal sheath cells expressing GFP::UNC-108. Two apoptotic germ cells engulfed by sheath cells are shown. Red arrow, appearance of GFP::UNC-108 on the phagosomal surface (dark green); black arrows, diffuse GFP::UNC-108 in the sheath cell cytosol (light green) separated by germ cell nuclei. (F and G) Total fluorescence intensity of GFP::UNC-108 on the phagosomal surface in embryos (F) or germ line (G) at different time points as shown in A–D was measured, and the normalized fluorescence intensity is shown. Enrichment of GFP::UNC-108 on the phagosomal surface was seen in wild type but not gop-1(tm5384) mutants.

Loss of gop-1 affects phagosomal recruitment of UNC-108. (A and B) Time-lapse images of somatic cell corpses in wild type (WT; A) and gop-1(tm5384) (B) expressing mCherry::MTM-1(C378S) and GFP::UNC-108. 0 min, time point when MTM-1(C378S) was first detected around the cell corpse. The time period until the cell corpse was surrounded by MTM-1(C378S) was defined as the pseudopod extension stage. Quantification of pseudopod extension and the phagosomal duration of MTM-1 and UNC-108 is shown at bottom (mean ± SD). n, number of phagosomes quantified. Yellow arrowheads, MTM-1 release from phagosomes; white arrowhead, recruitment of GFP::UNC-108 to the phagosomal membrane. (C and D) Time-lapse images of germ cell corpses in wild type (WT; C) and gop-1(tm5384) (D) expressing GFP::UNC-108 and mCherry::H2B (Histone 2B). 0 min, time point when phagosomal degradation of the apoptotic cell, indicated by diffuse H2B (white arrowheads), was observed. In wild type, UNC-108 was enriched on the phagosomal membrane 36 min before diffuse H2B was observed, whereas it was absent from phagosomes in gop-1(tm5384). Percentage of phagosomes with the representative pattern is shown in parentheses. n, number of phagosomes that were followed and quantified. In A–D, pink arrowheads indicate the first time point when the intensity of GFP::UNC-108 on the phagosomal surface (white dashed line) was measured. Bars, 5 µm. (E) Cartoon showing a cross section of a pair of gonadal sheath cells expressing GFP::UNC-108. Two apoptotic germ cells engulfed by sheath cells are shown. Red arrow, appearance of GFP::UNC-108 on the phagosomal surface (dark green); black arrows, diffuse GFP::UNC-108 in the sheath cell cytosol (light green) separated by germ cell nuclei. (F and G) Total fluorescence intensity of GFP::UNC-108 on the phagosomal surface in embryos (F) or germ line (G) at different time points as shown in A–D was measured, and the normalized fluorescence intensity is shown. Enrichment of GFP::UNC-108 on the phagosomal surface was seen in wild type but not gop-1(tm5384) mutants.

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