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Fig. 1. After exposure to X-rays, cellularized embryos delay metaphase-anaphase transition by a mei-41-dependent mechanism. (A) Irradiation delayed the metaphase-anaphase transition. Montages of two mitotic cells from the dorsal ectoderm of embryos expressing a GFP-Histone H2Av transgene are shown. Embryos were irradiated with 0 (–IR) or 8.3 Gy (+IR) of X-rays and irradiated embryos were rested for 40 minutes to allow cells to enter mitosis, prior to analysis. Frames were taken every 30 seconds for up to 10 minutes. Arrowheads mark the beginning of metaphase and arrows mark the end of metaphase. (B) The length of metaphase specifically lengthens following irradiation in the 14th, 15th and 16th cellular cycles. Embryos were collected for 30 minutes and aged 130 minutes to reach cycle 14, 240 minutes to reach cycle 15 or 330 minutes to reach cycle 16. Embryos were irradiated with 0 (–IR) or 8.3 Gy (+IR) of X-rays and analyzed 0 minutes (–IR) or 40-60 minutes (+IR) later. For each time point, at least seven nuclei from at least three different embryos were analyzed. Asterisks denote statistically significant data with P<0.0001. P values were generated from the comparison with the non-irradiated control for each cycle (i.e. Cyc14 +IR versus Cyc14–IR). (C) mei-41 is required to delay the metaphase-anaphase transition after irradiation. Wild type (Wt) and mei-41^D12 mutant embryos in cycle 16 (330-360 minutes after egg deposition, AED) were irradiated with 0 (–IR) or 8.3Gy (+IR) of X-rays and analyzed 0 minutes (–IR) or 40 minutes (+IR) later. For each time point at least 10 nuclei from four different embryos were analyzed. Asterisks denote statistically significant data with P<0.0001.

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