Translational inhibition ameliorates the Mybx1 phenotype. (A) Western blot of Ybx1, GFP-Eif4ebp1-TTAA and Gapdh protein expression in WT, Mybx1 and Mybx1+TTAA embryos. (B) Bright-field images showing representative Mybx1 and Mybx1+TTAA embryos with chorions after fertilization. (C) Chorion elevations of individual 0.5 hpf fertilized eggs from a single Zybx1 or Zybx1+TTAA female. Each dot or circle represents one egg's value. ***P<0.001; Welch's t-test. (D) Average chorion elevations of fertilized eggs from different females. Six females of each group, 50-100 eggs per female, were analyzed. Each circle represents the average of all observed eggs from a single female. *P<0.05; Welch's t-test. (E) Coomassie staining of major yolk proteins of ybx1 mutant and ybx1+TTAA stage-V eggs. HYP/LYP ratios were calculated to display yolk protein cleavage levels. (F) Major yolk protein cleavage in oocytes before and after in vitro maturation. (G) Bright-field images of Mybx1 and Mybx1+TTAA gastrulas. (H) The percentage of epiboly restored Mybx1+TTAA embryos from different females. Epiboly initiation is extremely rare in Mybx1 mutants. *P<0.05; n=6; Welch's t-test. (I,J) qRT-PCR detection of partially restored maternal mRNA clearance (I) and zygotic gene transcription (J) in Mybx1+TTAA embryos, compared with Mybx1 mutants. WT results are also presented. *P<0.05, **P<0.01; n=3; Student's t-test. Scale bars: 200 μm.
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