Fig 5. Extensive transcriptome changes, and high correlation with Dicer KO oocytes, in Ago2 ^ADH oocytes.

Oocytes from Ago2 ^fl/fl, Ago2 ^ADH, Ago2 null, Dicer WT, and Dicer KO females were subjected to RNA-seq. A) Comparison of transcripts up-regulated (↑) or down-regulated (↓) in Ago2 ^ADH vs. Ago2 ^fl/fl oocytes (blue circles) with those up-regulated (↑) or down-regulated (↓) in Dicer KO vs. Dicer WT oocytes (green circles). Mis-regulated transcripts were identified using an FDR of 0.01. The overlapping transcripts are shown in red. *p< 2.2e-16, Chi-square test. B) The majority of genes that produce endo-siRNAs in oocytes are up-regulated in the absence of AGO2 catalytic activity. Transcript levels in our RNA-seq dataset were compared in Ago2 ^ADH vs. Ago2 ^fl/fl oocytes for the 20 genes that produce the largest number of endo-siRNAs [9] and fold-changes were calculated. C) Validation of RNA-seq data by qRT-PCR. The relative abundance of 11 selected transcripts [6 up-regulated (↑), 3 unchanged, and 2 down-regulated (↓)] in our RNA-seq dataset when comparing Ago2 ^ADH vs. Ago2 ^fl/fl oocytes) was determined in oocytes of the different Ago2 genotypes by qRT-PCR. Transcript levels in Ago2 ^fl/fl oocytes were set as 1. Data are expressed as the mean ± SEM of 3 experiments. *p< 0.05 vs. Ago2 ^fl/fl; two-way ANOVA, followed by Bonferroni post-test.