Activation of Chk2 in the mof heterozygous embryos. (A) Total RNA was isolated from embryos of syncytial cycles of 10–13 from control yw67c23 and mof1 embyros and semi-quantitative RT-PCR was carried out using primers against ATM, ATR, Chk1 and p53 to study their expression pattern. (B) Semi-quantitative RT-PCR was carried out using chk2 primers in yw67c23 and mof1 embryos (syncytial cycles 10–13). We observed 4-fold increase in the level of chk2 in mof1 embryos compared to control indicating activation of chk2. (B’) Depletion of MOF by RNAi was performed in S2 cells (Drosophila Schneider cells) by incubation with dsRNA against mof. The transfection was conducted for 72 h time period. Total RNA isolated was subjected to RT-PCR analysis against Chk2 and mof. Here dsRNA against GFP was used as control (control RNAi). The depletion of mof leads to an increase in the levels of Chk2 mRNA. (C) Early embryos from mof1 and mof1/+; mnkp6/+ females were collected and stained with DNA dye PI to study the severity of nuclear fallout. The high severity of nuclear fallout in mof1 embryos was drastically reduced in the presence of mnkp6(chk2) mutation and is graphically represented. Statistical significance was assessed using student t-test. *** indicates P<0.001, ** indicates P<0.01, * indicates p<0.05. Bar indicates 10 μm scale.