MED13 knockdown impairs transcription of genes required for the OET, including cell
cycle and chromatin-remodeling proteins. One-cell stage embryos were microinjected with
the indicated morpholino, then cultured to the late 2C stage. RNA sequencing was
performed on independent pools of 40 embryos per group. (A) Unsupervised hierarchical
clustering of gene expression differences in Scr-MO (Scr)- and Med13-MO (M13)-injected
embryos. (B) Numbers of altered transcripts of the indicated types in Med13-MO-injected
embryos relative to Scr-MO-injected embryos. (C) Highly significant GO biological
categories of genes downregulated in Med13-knockdown embryos. Numbers to right of bars
indicate number of genes in category. (D) Selected GO cell cycle genes significantly
altered in Med13-knockdown embryos. (E) Selected GO chromatin modification genes
significantly altered in Med13-knockdown embryos. (F) Percentage of embryos to reach the
indicated preimplantation embryo stages following microinjection at the 1C stage with
the either Scr-MO, Med13-MO, or Med13-MO and both Smarca4 and
Smarcc1 mRNAs. N = 3, 8–12 1C embryos/group for each replicate;
*P ≤ 0.05 compared to Scr-MO control at the same time point,
#P ≤ 0.05 compared to MED13-MO at the same
time point, ANOVA with Holm-Sidak multiple comparisons test. Graph shows mean ± s.e.m.
(G) Graph showing total number of chromosomal regions either up- or downregulated for
each of the indicated repetitive element classes.