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. 2011 Apr 8;6(4):e18325. doi: 10.1371/journal.pone.0018325

Figure 2. LST-4 is recruited to the early phagosome where it colocalizes with DYN-1 and RAB-5.

Figure 2

(A–G) DIC micrographs (A–D) or epifluorescence pictures (A′–C′, E–G) of C. elegans germ lines. Arrowheads indicate early apoptotic germ cells or protein localized around apoptotic germ cells. LST-4 is recruited around the apoptotic cell during internalization (A′, E, G arrowhead) and highlights early, SYTO negative corpses (F). SYTO, like Acridine Orange, preferentially stains late-stage, internalized apoptotic cells. When the internalization process is disrupted, as in ced-1(e1735) (B, B′) and ced-12(k149) mutants (C, C′), LST-4 localization around apoptotic germ cells is lost (B′, C′ arrowheads). (H–O) DIC micrographs (H, L) or epifluorescence pictures (I–K, M–O) of C. elegans germ lines. LST-4::CFP (I, M) extensively colocalizes with DYN-1::YFP (J, K), but not with YFP::RAB-7 (N, O). (P) Localization of LST-4::YFP in different genetic backgrounds. Germ cell corpses and LST-4 positive phagosomes were quantified as described in materials and methods. Data shown are mean ± s.d. n>15 animals for each condition. Size bars, 10 µm. (R, S) Colocalization index of LST-4::CFP with YFP::actin, YFP::DYN-1, YFP::RAB-5 and YFP::RAB-7. Data shown are mean ± s.e.m. n>50 halos for each genotype. Determination of the colocalization index is described in materials and methods.