Chordoma epithelial characteristics in single cells and clusters. Human chordoma tissue (42 resections, 23 patients). (A) U-CH1 single cells, F-actin (green), and cytokeratin 8 and 18 (red) expression. (B) U-CH1 single cells containing actin (green), α6 integrin (red), and DNA (blue). (C) U-CH1 clusters containing actin (green), α6 integrin (red), and nuclear DNA (blue). (D and E) Human chordoma specimen stained for α6 integrin (brown). (F) The percentage of cells in the population contained within cohesive clusters under normal coating conditions (gelatin) or grown on immobilized HYD1 (HYD1), a laminin ligand mimetic. Flow cytometry profiles of (G) brachyury and (H) YAP. (I) Expression and mean peak fluorescence values for surface expression of α6 integrin (CD49f), α3 integrin (CD49c), β1 integrin (CD29), β4 integrin (CD104), and MUC1 (CD44) in UCH-1 (solid black) or CHOR1 (striped) cells.