Figure 6. Zygotic depletion of Zmym2 arrests development at 2C-stage embryos.

(A and F) Morphology of embryos after zygotic injection with miR-344-3p mimics (mimics) and negative control (miNC) (A) or siRNA against Zmym2 (siZmym2) and non-targeting siRNA (siNC) (F) at each indicated developmental stage. The blue squares indicate aberrantly developed embryos. Scale bars, 50 μm.

(B and G) Statistics for the development efficiency (portion of normal embryos) at each developmental stage in Figure 6A (B) or Figure 6F (G). Data are presented as average percentage ± SD. P-value is from two-way ANOVA test; n.s. non-significant; *** P<0.001.

(C-E) RT-qPCR of MERVL/Zscan4 (C) and Zmym2/Lsd1 expression (D) in embryos with miR-344-3p mimics (mimics) or negative control (miNC) treatments, collected at the 8-cell stage or at the blastocyst stage (E). Data are presented as average ± SD. P-value is from T-test, ** P<0.01.

(H) RT-qPCR analysis of Zmym2 and MERVL expression in embryos with siZmym2 or siNC. Embryos were collected at the 8-cell and blastocyst stages, and siZmym2-treated embryos with normal or arrested morphology were collected separately. Data are presented as average ± SD. P-value is from T-test, * P<0.05, ** P<0.01.

(I) PCA of RNA-seq data of embryos with miR-344 mimics, siZmym2, and non-target control (NC) injections from this study, and of normal embryos at different embryonic stages from (Liu et al., 2012). Each sample represents a pool of embryos (~10) with identical treatment and developmental stage. Shapes of samples indicate the treatments, and colors of samples indicate the stage for RNA collection. The curved arrow indicates development progression from zygote to blastocyst. Two pools of embryos (clear triangles) are developmentally arrested upon siZmym2 treatment. RNA is collected at 8-cell/early morula stage.