Fig. 5
The undead B.al/rapaav cell in engulfment mutants fails to display characteristics of dead cells. a–c A living B.al/rapaav cell (cell boundary indicated by dotted line) in the early fourth larval stage before cell death is complete (a) lacks refractility in the nucleus, (b) has P egl-1 ::4xNLS::GFP in the nucleus but not in the rest of the cell and (c) has P evl-20 ::mCherry::PH localized to the membrane. d–f A B.al/rapaav cell corpse has (d) refractility throughout the cell, (e) P egl-1 ::4xNLS::GFP throughout the cell, and (f) P evl-20 ::mCherry::PH throughout the cell. g–i An undead B.al/rapaav cell in the late fourth larval stage of an engulfment-defective mutant (g) lacks refractility in the nucleus, (h) has P egl-1 ::4xNLS::GFP in the nucleus but not the rest of the cell, and (i) has P evl-20 ::mCherry::PH localized to the membrane. j–l An undead B.al/rapaav cell in the late fourth larval stage of a suicide-defective mutant (j) lacks refractility, (k) has P egl-1 ::4xNLS::GFP in the nucleus but not the rest of the cell, and (l) has P evl-20 ::mCherry::PH localized to the membrane. Animals were of genotypes nIs343; him-5(e1467ts) unc-76; nIs349; nEx2344 (a–f), nIs343; ced-10 him-8; nIs735 (g–i), or nIs343; ced-3 him-8; nIs735 (j–l). Scale bars: 10 μm. m A B.al/rapaav cell (arrow) in the early fourth larval stage has diffuse chromatin similar to neighboring living cells. n A dying or dead B.al/rapaav cell in the mid-fourth larval stage has condensed chromatin. o An undead B.al/rapaav cell in the late fourth larval stage of an engulfment-defective mutant has diffuse chromatin similar to neighboring living cells. p An undead B.al/rapaav cell in a suicide-defective mutant has diffuse chromatin similar to neighboring living cells. Scale bars: 10 μm. Animals were of the genotypes nIs343; him-5(e1490); nIs349 (m, n), ced-12 ced-1; nIs343; him-8 (o), or nIs343; ced-3 him-8 (p). a–l depict different animals than in (m–p), since the fixation required for visualizing DAPI staining precludes the use of DIC optics