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. Author manuscript; available in PMC: 2010 Jun 16.
Published in final edited form as: Biochemistry. 2009 Jun 16;48(23):5396–5404. doi: 10.1021/bi900253d

Figure 4. AP site cleavage is inefficient in eggs and less efficient in pre-hatching embryos than at later times.

Figure 4

A. Extracts (2 μg protein) from eggs or adult fish were incubated for the indicated time in the presence of 5 mM Mg2+ with 5′-end labeled substrate that had been treated with Ung to remove uracil for varying time intervals (21). Reactions were stopped by addition of EDTA, and phenol extracted. Substrate and product were resolved by gel electrophoresis. AP site cleavage was linear for ∼2 min under these conditions. The rate of AP site cleavage was then examined in extracts (2 μg protein) obtained from eggs (●) or adult fish (■). B. AP site cleavage by eggs, embryos and adult fish (2 μg protein) over a 1 min interval. *: 3.5 hpf embryos in which ZAP1 has been reduced by 74-90 % by microinjection of 0.5 mM TS-MO at the 2-4 cell stage; **: extracts from 6.5 hpf embryos in which ZAP1 has been reduced by 40-56 % by microinjection of 0.2 mM TS-MO. These data are the average of three experiments +/- SE.