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. 2022 Jul 7;11:e78829. doi: 10.7554/eLife.78829

Figure 4. Smooth chorion-specific cytotrophoblasts (SC-CTBs) express a distinct epidermal transcriptional program.

(a) Heatmap of gene ontology analysis adjusted p-values. Dark red corresponds to the lowest p-values and white represents p-values greater than 0.0005. Ontology categories are organized by hierarchical clustering along the y-axis. Marker genes for each cluster were used as inputs for the analysis. A subset of genes in selected categories are listed at the right. Categories and genes enriched in CTB 1 or CTB 4 are in black or blue, respectively. (b) Representative immunofluorescence co-localization of non-phosphorylated CTNNB1 and KI67 in the VC (left) and SC (right). (c) Quantification of the percent of np-CTNNB1 cells with KI67 expression in each region in three placental samples. Each dot represents the percentage in one field of view (at least seven per region per sample) as an estimate of mitotic cells per population. Percentages for the villous chorion (VC) region are shown in black and the SC region in blue. (d) Immunofluorescence co-localization of CDH1 and KLF4 protein in the VC (left) or SC (right). Arrowheads denote CDH1+/KLF4+ cells. (e) Immunofluorescence co-localization of KRT14 and KRT6 protein in the VC (left) or SC (right). The outline of the anchoring villi (AV) is denoted by the white dashed line. For all images, nuclei were visualized by DAPI stain; scale bar = 100 μm. Abbreviations: FV = floating villi; SC = smooth chorion epithelium; Amn. = amnion; Dec. = decidua; GV = ghost villi.

Figure 4.

Figure 4—figure supplement 1. Complete cytotrophoblast (CTB) gene ontology analysis.

Figure 4—figure supplement 1.

The unabridged gene ontology results from each CTB cluster, displayed as adjusted p-values. Dark red corresponds to the lowest p-values and white represents p-values greater than 0.0005. Ontology categories are organized by hierarchical clustering along the y-axis. Marker genes for each cluster were used as inputs for the analysis.
Figure 4—figure supplement 2. Similarities and differences between cytotrophoblast (CTB) 1 in villous chorion (VC) and SC.

Figure 4—figure supplement 2.

(a) Immunofluorescence co-localization of CDH1 and non-phosphorylated CTNNB1 in the VC (left) and SC (right), showing identical domains of expression in both regions. For all images, nuclei were visualized by DAPI stain; scale bar = 100 μm. Abbreviations: FV = floating villi; MB = maternal blood space; SC = smooth chorion epithelium; Dec. = decidua. (b) Gene ontology analysis for CTB 1 genes common to VC and SC, genes enriched in VC, and genes enriched in SC. (c) Violin plots of genes within the gene ontology categories indicated in (b) for common categories (top), VC enriched categories (middle), and SC enriched categories (bottom).
Figure 4—figure supplement 3. Cytokeratin expression in villous (VC) and smooth chorion (SC) trophoblast.

Figure 4—figure supplement 3.

(a) Violin plots of cytokeratins expressed in trophoblast in both VC and SC regions by cluster and region of origin. (b) Violin plots of cytokeratins expressed in SC trophoblast by cluster and region of origin.
Figure 4—figure supplement 4. IFITM3 expression in cytotrophoblast (CTB) populations.

Figure 4—figure supplement 4.

Violin plot of IFITM3 expression in trophoblast clusters by region, showing increased expression in smooth chorion (SC) compared to villous chorion (VC).