Figure 4.
CEP-1 is expressed and induces EGL-1 transcription in response to DNA damage or unrepaired recombination intermediates in both hermaphrodite and male germ lines. (A) Immunolocalization of CEP-1 (red) counterstained with DAPI (blue) in pachytene nuclei from wild-type and cep-1(gk138) hermaphrodite and male germ lines. Scale bars, 5μm. (B) egl-1 transcript levels are induced following DNA damage or in the presence of unrepaired meiotic DSBs in hermaphrodites and males. Relative egl-1 transcript levels as determined by qRT-PCR following IR and/or in mutant germ lines. RNA was extracted from wild-type and cep-1 hermaphrodites and males 24h following treatment with IR (120 Gy) and 24h post-L4 for zim-1 hermaphrodites and males. (C) Transcriptional induction of egl-1 in zim-1 hermaphrodite and male germ cells is dependent on cep-1 and chk-1. Relative mRNA levels were determined by qRT-PCR in zim-1 L4440, zim-1;cep-1(RNAi) and zim-1;chk-1(RNAi) hermaphrodites (left) and males (right) 24h post-L4. Data shown for B and C are representative of three independent extractions and qRT-PCR reactions; error bars indicate standard deviation of 3 individual replicas of each sample.