Fig. 8: Functional investigation of SNAI1 on EVT cell differentiation in CT27 cells.

a RT-qPCR measurement of SNAI1 (n = 5 biologically independent replicates per group) normalized to B2M in extravillous trophoblast (EVT) cells differentiated from stem state cells transduced with lentivirus containing a control shRNA (shC; blue) or SNAI1-specific shRNA (shSNAI1; gray). Data were analyzed by unpaired t-test and are presented as mean values ± standard deviation (SD) and **p = 0.0054. b SNAI1 (29 kilodaltons (kDa)) and GAPDH (37 kDa) protein in EVT cells from shC control or shSNAI1 treated cells measured by western blot. c Phase contrast images of shC control or shSNAI1 EVT cells following eight days of differentiation. Scale bars represent 500 μm. d Heat map based on scaled, normalized counts of selected transcripts generated from RNA-Seq in shC (control, Ctrl; orange) or shSNAI1 (knockdown, KD; blue) transduced cells (n = 3 biologically independent replicates per group) using a red-blue diverging scale. Transcripts are clustered into three groups including EVT-specific (pink), EVT interferon (IFN) responsive (purple), or stem state-specific (green). e Hi-C, RNA-Seq, ATAC-Seq, and H3K4me3 and H3K27ac ChIP-Seq assessments performed in CT27 cells differentiated into EVT cells (top panel) or maintained in the stem state (bottom panel). Identified regulatory elements near SNAI1 are highlighted in orange and overlap Hi-C loops (red, both loop anchors in view; blue, loop anchors out of view), open chromatin by ATAC-Seq (counts per million mapped reads, y-axis), and promoter or enhancer mark by H3K4me3, or H3K27ac ChIP-Seq, respectively (counts per million mapped reads, y-axis). Expression level is shown by RNA-Seq (transcripts per million, y-axis). All datasets are shown in individual tracks. Super-enhancers (SE, red) and differentially bound regions (DB, blue) are specific to the EVT cell state. Source data are provided as a Source Data file.