Aldefluor expression is enhanced in HM-LLC but not in TC-1 spheres compared to adherent cells.

<p>(<b>A–B</b>) Representative plots showing Aldefluor staining (blue) and DEAB control staining (red) for HM-LLC, LM-LLC and TC-1 adherent cells compared to passage 2, day 1 and day 7 sphere-derived cells. Aldefluor high expressing cells (rectangular box) are significantly greater in (<b>C</b>) frequency and (<b>D</b>) mean fluorescent intensity (MFI) of Aldefluor within HM-LLC spheres compared to LM-LLC or HM-LLC adherent cells, and HM-LLC adherent cells express more Aldefluor than LM-LLC adherent cells; frequency (<b>C</b>) *P≤0.001; MFI (<b>D</b>) *P≤0.002. Compared to DEAB staining, Aldefluor positivity (non-rectangular box) was found to be significantly higher in TC-1 adherent cells compared to spheres (<b>E</b>); *P≤0.004. MFI of Aldefluor within the TC-1 groups (<b>F</b>) was not found to be significantly different. No Aldefluor high population was noted for TC-1 spheres or adherent cells. Plots represent combined data from two biological repeats with three technical replicates each. Bars: ±SEM.