Wdr74 is required for blastocyst formation.

<p><b>A.</b> Quantitative RT-PCR analysis of endogenous <i>Wdr74</i> mRNA during preimplantation development. <b>B.</b> RT-PCR with <i>Wdr74</i> intron-spanning primers confirms relative abundance of transcripts observed by qRT-PCR. <b>C–H.</b> Microinjected and cultured embryos photographed at 36, 60, and 84 hours post fertilization. Control dsGFP-injected embryos show normal development and form blastocysts by 84 hpf (C–E). dsWdr74 injected embryos develop normally to the morula stage (F–G) but fail to make blastocysts (H). <b>I.</b> Quantification of percent 2 cell embryos that develop to the blastocyst stage by 84 hpf (# blastocysts/# 2-cell ×100). <b>J.</b> qRT-PCR of Wdr74 transcripts indicates robust RNAi mediated knockdown due to microinjection of dsWdr74. <b>K.</b> Immunofluorescence of Wdr74 in morula stage dsGFP embryos shows nuclear localization; which is drastically reduced in dsWdr74 embryos of the same stage (L). hpf, hours post fertilization. Results of student T-test shown, error bars represent standard deviation. All data shown normalized to embryo equivalents; MII, Metaphase II oocyte. Scale bar in F representative for C–H. K′ and L′ show DAPI signal (DNA) from the same embryos shown in K and L, respectively.