# Placental Transcriptome Adapations to Maternal Nutrient Restriction in Sheep
## 2 Results
### Over-Representation Analysis of GO: Molecular Functions  
Genes that were differentially expressed between the three treatment groups on day 70 of pregnancy were further analyzed to determine changes in Gene Ontology (GO) molecular functions (Figure 2). Only functions with > 1 gene involved and an FDR < 0.05 are shown. While the number of DEGs was fairly low in each comparison at day 70, there were still nine functions that were differentially enriched between NR SGA and 100% NRC placentomes, including terms such as MHC class II protein binding, cytokine binding, and transmembrane signaling receptor binding (Figure 2A). MHC class II receptor activity was the only function noted in NR NonSGA compared to 100% NRC placentomes, and the heat map shows that two of the three involved genes (_major histocompatibility complex_, _class II_, _DQ alpha 2 (HLA-DQA2_) and _major histocompatibility complex_, _class II_, _DQ beta 2 (HLA-DQB2_)) were downregulated in NR NonSGA compared to 100% NRC placentomes, while _HLA-DOB_ was upregulated in NR NonSGA placentomes (Figure 2B). In NR SGA compared to NR NonSGA placentomes there were eight functions noted that included  
\begin{table}
\begin{tabular}{c c c c c c c c} \hline \hline \multicolumn{3}{c}{**NR SGA 1 vs. 100\% NRC**} & \multicolumn{3}{c}{**NR NonSGA vs. 100\% NRC**} & \multicolumn{3}{c}{**NR SGA vs. NR NonSGA Cont. **} \\ \hline
**Gene** & **Log2 FC 2** & **FDR 3** & **Gene** & **Log2 FC** & **FDR** & **Gene** & **Log2 FC** & **FDR** \\ \hline _CLRN2_ & \(-3.82\) & 0.04 & _CYP3A5_ & \(-3.68\) & 0.04 & _LPCAT4_ & \(-0.48\) & 0.02 \\ _PHWL1_ & \(-3.34\) & 0.09 & _CYP3A24_ & \(-3.36\) &  
terms such as receptor ligand activity, organic hydroxyl compound transporter activity, and lipid transporter activity (Figure 2C). Signaling receptor binding and cytokine activity were also functions of note identified in NR SGA versus NR NonSGA placentomes, and as they were also identified at day 135, the expression for the genes involved in those functions is shown in a heat map in Figure 2D. Genes that are upregulated in NR SGA placentomes include _family with sequence similarity 3 member C_ (_FAM3C_), _S100 calcium-binding protein B_ (_S100B_), and _osteoprotegerin_ (_TNFRSF11B_), while _apolipoprotein E_ (_APOE_), _HLA-DOB_, _junctional adhesion molecule 2_ (_JAM2_), _MSTN_, and _Wnt family member 9B_ (_WNT9B_) were downregulated. Molecular functions that were differentially enriched between treatment groups at day 135 are shown in Figure 3. There were seven functions altered between NR SGA and 100% NRC placentomes, which included terms such as signaling receptor binding, molecular  
Figure 2: GO molecular function analyses of differentially expressed genes (DEGs) in placentomes at day 70 of pregnancy. Molecular functions differentially regulated in NR SGA compared to 100% NRC placentomes (**A**). The molecular function differentially regulated in NR NonSGA compared to 100% NRC placentomes, with a heat map of the three involved genes (**B**). Molecular functions differentially regulated in NR SGA compared to NR NonSGA placentomes (**C**), \({}^{\star}\) with a heat map of genes involved in “signaling receptor binding” and “ cytokine activity” (**D**). The heat maps correspond to one sample for each column and one gene for each row. All depicted GO molecular functions were significant at FDR < 0.05. Numbers in parentheses indicate the number of DEGs involved in the function. transducer activity, and polysaccharide binding (Figure 2A). Since only seven genes were differentially expressed between NR NonSGA and 100% NRC placentomes, only functions involving one or two genes were identified; those functions include hydroxylase activity for multiple molecules and oxidoreductase activity (Figure 3B). There were 15 functions with >1 gene altered in NR SGA compared to NR NonSGA placentomes, with terms such as CXCR chemokine receptor binding, amyloid-beta binding, and lipid kinase activity (Figure 3C).