fig1

Figure 1. Characterization of cancer stem-like properties of UP-LN1 cells. (a) The side-population method was employed to compare and analyze the percentage of potential stem cell-like cells in UP-LN1 cells. With the absence and presence of verapamil, the percentage of side-population cells was calculated. In the upper frame, based on representative results of one experiment, parental (P) contained an intermediate of 2.93% side-population cells, adherent (A) cells contained the least among the three groups at 1.07%, and floating (F) cells contained the highest percentage at 4.20%. The quantitative results shown in the lower frame reveal that the percentage of side-population in F cells is significantly higher than that in A and P cells with P < 0.01 and P < 0.05, respectively; (b) to support our side-population data, an embryonic stem cell-specific fluorescent dye CDy1 was used to stain UP-LN1 cells. The red fluorescent signal was strongly associated with F cells as compared to A cells (arrowheads). Notably, red fluorescence was signifi cantly stronger in F-cell aggregates (arrows); (c) when examined by Western blot analysis, F cells were found to express a significantly higher level of stemness genes (including Nanog, c-Myc, Oct4, and Sox2) than P and A cells. Note that the relative densities in the expression of each stemness gene among F, P, and A cells are also shown by fold difference in this figure