Fig. 4

Effect of FOXO1 overexpression on FOXL2^C134W – modified genes. a Venn diagram showing the number of differential genes (DESeq, q < 0.05) identified in FOXL2^C134W vs FOXL2^WT and FOXL2^C134W + FOXO1 vs FOXL2^WT. b Scatter plot of the effect sizes (log2-fold change) of DEGs between FOXL2^C134W and FOXL2^WT, with (y-axis) and without (x-axis) the addition of FOXO1. Genes that were differentially expressed also when adding FOXO1^3A are shown in black (n = 130), while genes that were differentially expressed only without FOXO1^3A addition are shown in purple (n = 587). The slope (ß) of the linear relationship between the two effect sizes is shown on top of the plot. c Venn diagram showing the number of differential genes (DESeq, q < 0.05) identified in FOXL2^C134W vs FOXL2^WT and FOXL2^C134W + FOXO1 vs FOXL2^C134W. d Heatmap with gene clustering of the 39 DEGs in both FOXL2^C134W vs FOXL2^WT and FOXL2^C134W + FOXO1 vs FOXL2^C134W comparisons. For each gene, expression values were z-score normalized across samples. Genes are annotated for presence or absence of a FOXL2^C134W ChIP-seq binding site in TGFβ-treated HGrC1 (ChIP1) [37] and a FOXL2^C134W ChIP-seq binding site in a SVOG3e cell line (ChIP 2) [28]. e Metascape GO analysis of the 39 DEGs in both FOXL2^C134W vs FOXL2^WT and FOXL2^C134W + FOXO1 vs FOXL2^C134W comparisons. The red line indicates p-value cutoff for p < 0.05. GO terms associated with expression of FOXL2 and/or SMAD3 were removed from the results (see Methods). f qPCR validation of the RNAseq expression data in FOXO1 modulated genes (BAZ2A, SMARCA4, COL4A2, and HSPG2). Graphs show fold change means ± s.e.m relative expression to FOXL^WT following normalization to the housekeeping gene. Data were analyzed using the two-tailed unpaired t-test (*p < 0.05, **p < 0.01, ***p < 0.001)