Fig. 1From: Angiopoietin-1 receptor Tie2 distinguishes multipotent differentiation capability in bovine coccygeal nucleus pulposus cellsSorting and gating strategies for Tie2+ cells from a whole NPC population. The NPC suspension after enzymatic digestion was colabeled with the Tie2 antibody and PI and sorted for the Tie2 marker. a, b Two examples show gating of the whole cell population for forward and side scatter (FSC and SSC, P1; left panel). It is important to mention that primary NPC after enzymatic digestion contain tissue fragments, granules of dead cells, and debris, which are removed by a selective gating for FSC and SSC (left panel, b). In addition doublets are excluded by a FSC-H versus FSC-A gating (middle panel, b). Proper gating for Tie2 is shown for the two examples and was performed by a negative selection of cells in isotype-matched control with less than 0.1 % (top right panel, P3) and setting the gate at the left for the Tie2– cells (P2). The same gating was then applied for the specific Tie2 staining and by excluding PI-positive cells. P1 whole NPC population, P2 Tie2– cell population, P3 Tie2+ cell population, PI propidium iodide, Tie2 angiopoietin-1 receptorBack to article page