To examine
lim-4 expression during the embryogenesis, the eggs were stained at various developing stages with anti-myc antibodies. A GFP reporter was used for the GATA gene
elt-2, which is specifically expressed in the gut lineage, to stage the embryos. MYC::LIM-4 immunoreactivity is first detected in two cells approximately 160 min post-fertilization at 8 gut nuclei stage. The two MYC::LIM-4-staining cells was identified as the neuroblasts ABalpppppa and ABpraaappa that respectively give rise to left/right ADF and AWB neurons, based on triple labeling of the embryos with antibodies raised against the LIN-26 protein, which is expressed in the nuclei of the hypodermal cells that border the ADF/AWB mother cells. Like
lim-4::gfp, the MYC::LIM-4 immunoreactivity is not detected in ADF at any stage of larvae or adults, but it is observed in AWB and several other neurons in the head throughout life (N > 200). It is difficult to determine if LIM-4 is excluded from ADF or it is present there transiently.