HCT116 FUT9 knockdown and matching control cells were seeded in ultra‐low attachment plates and cultured for 1 week. The resulting tumorspheres were collected, dissociated, and the total number of cells counted. The graph represents the mean ± s.e. of two independent replicates normalized to the number of control cells. Each replicate represented tumorsphere cells collected from 24 independent wells. Representative images are shown. Scale bar, 1,000 μm.
FUT9‐overexpressing and control cells were cultured as tumorspheres and analyzed as in (A). Two independent replicates and representative pictures are depicted. The graph represents the mean ± s.e. of two independent replicates normalized to the number of control cells. Representative images are shown. Scale bar, 1,000 μm.
CD44 expression in FUT9 knockdowns (in red) and shRFP control (in blue) in HCT116 cells was assessed using anti‐CD44 and flow cytometry, and representative histograms were overlayed (second panel). Isotype controls were also plotted and overlayed (first panel). Median fluorescent intensity (MFI) values derived from the software are plotted as bar chart. The graph represents the mean ± s.e. of two independent replicates.
HCT116 FUT9 knockdown or control cells were injected subcutaneously into the right flank of NOD/SCID mice and monitored for tumor formation. Each tumor was measured using calipers, and the mean volume for the FUT9 knockdown and control tumors was graphed (first panel). The graph represents two independent experiments with a minimum of 11 mice analyzed per experimental condition. Mean tumor volumes ± s.d. are shown. Upon experiment termination, tumors were extracted, weighed, and the mean tumor weights ± s.d. are shown in the second panel.
A schematic showing the abundance of FUT9‐positive cells over the course of colon cancer development.
‐test.
