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Table 2 Overview of the aphidicolin, actinomycin D and puromycin treatment experiments on Mnemiopsis leidyi embryos

From: A cleavage clock regulates features of lineage-specific differentiation in the development of a basal branching metazoan, the ctenophore Mnemiopsis leidyi

Developmental stage Treatment Observation Conclusion
2-cell stage Arrest of DNA synthesis for 2 h with aphidicolin, 2 μg/ml No combs formed (0/30) DNA synthesis is essential for comb formation.
16-cell stage
32-cell stage
60-cell stage 27/30 (90%) formed combs, comb formation was delayed by 2 h
4 hpf 29/30 (97%) formed combs, comb formation was delayed by 2 h
5 hpf 29/30 (97%) formed combs, comb formation was delayed by 2 h
60-cell stage and 4 hpf Arrest of DNA synthesis and cytokinesis with aphidicolin, 2 µg/ml, plus cytochalasin B, 1 to 2 μg/ml Immediately arrested, single nuclei observed, neither combs nor photoprotein were formed in any embryo (0/30) DNA synthesis is essential for comb formation even in cleavage-arrested embryos.
1- to 2-cell stage Blocking of transcription with actinomycin D, 10 μg/ml 0/30 gastrulated, development arrests at 32- to 60-cell stage Zygotic transcription begins around the 32- to 60-cell stage and the specification of the combs requires more than only maternal RNAs.
4- to 8-cell stage 0/30 gastrulated, development arrests at 32- to 60-cell stage
16- to 32-cell stage 10/30 (33%) gastrulate, 20/30 (67%) arrest at 32- to 60-cell stage
60-cell stage 28/30 (93%) gastrulate but no pharynx is formed, 2/30 (7%) do not gastrulate
5 hpf (post-gastrulation) No pharynx is formed
2-cell stage Arrest of translation with puromycin, 125 μg/ml, 25 μg/ml 0/30 formed combs Comb protein is translated prior to 6 hpf, comb formation requires protein that is translated in the embryo and not maternally loaded.
16-cell stage 0/30 formed combs
60-cell stage 0/30 formed combs
6 hpf (post-gastrulation) 32/30 (97%) formed combs