Fig. 1

a ESCs were infected with lentiviruses encoding shRNAs to Sall4 and Wapal. Cells with a high multiplicity of infection (MOI) were selected by addition of puromycin for 6 days. Images were taken by bright field (BF, ×10) or after staining for the pluripotency marker alkaline phosphatase (AP, ×10). b Changes in pluripotency markers (Nanog, Oct4, Sall4, and Rex1) and Wapal were measured by RT-qPCR. Fold change was calculated relatively to cells infected with the empty vector and plotted linearly on the y-axis. Error bars represent SEM of at least two experiments. Asterisk indicates statistically significant reductions compared to empty vector (p value <0.05). c Similar to b, but for differentiation markers Brachyury (mesoderm), Fgf5 (ectoderm), and Cdx2 (trophectoderm). a Log₁₀ scale for fold change is used because of strong induction of these markers. Asterisk indicates statistically significant increases compared to empty vector (p value <0.05). d Protein levels after depletion of Wapal or Sall4 for core cohesin subunits Rad21 and Smc3. GAPDH is shown as a loading control. Intervening, unrelated lanes were removed, but all samples were run on the same gel. e Propidium iodide (PI) staining was used to measure DNA levels in cells by flow cytometry. The percent of cells within each phase of the cell cycle and which exhibited abnormal DNA content (<2N or >4N) is shown. The only statistically significant difference was in the fraction within G0/G1 after depletion of Sall4 or Wapal as compared to empty vector (asterisk indicates p value <0.05)