Fig. S1. Molecular cloning of
tbc-2. (A) Cloning of
tbc-2. The top bar indicates the genetic map of the
tbc-2 region and the lower panels show the rescue of
tbc-2(
qx20). Non-transgenic and transgenic embryos at the 2-fold stage were scored for each construct. At least 15 animals (transgenic or non-transgenic) from each independent transgenic line were scored for all lines obtained, as indicated in parentheses. The
tbc-2 gene structure is shown, with filled boxes representing the exons and the interspersed lines indicating the introns. The arrow pointing away from the filled boxes shows the direction of
tbc-2 transcription. The mutation identified in the
qx20 mutant is marked and the gray bar below
tbc-2 transcript delineates the actual region of the
tbc-2 gene removed by the deletion mutant
tm2241, with the sizes of the deletion and insertion shown. (B) TBC-2 is expressed in the known engulfing cells. The expression of TBC-2::GFP driven by the
tbc-2 promoter was examined in wild-type animals. TBC-2::GFP was seen in several known engulfing cell types, such as hypodermal cells (a,b), pharyngeal muscle cells (c,d) and intestinal cells (e,f). TBC-2::GFP was also found to cluster around the cell corpse as indicated by the arrows in g and h. (C) GDP-locked RAB-5 failed to associate with phagosomes. DIC and fluorescence images of a wild-type embryo expressing GFP::RAB-5(S33N), a GDP-locked RAB-5, are shown. GFP::RAB-5(S33N) failed to cluster around cell corpses (indicated by arrows) and displayed a diffused cytoplasmic pattern. Scale bars: 5 µm.