Figure 3

MiR-21 was up-regulated in CD133+ PA1 cell populations. APC-conjugated CD133 antibody was used to enrich CSPCs by FACS sorting. The basal IgG-isotype-APC staining signal peaks are presented on the left-hand side and the CD133-APC staining signal peaks are presented on the right-hand side of the histogram. As indicated in P2 and P3, the 5% extremes of the staining signal within the spectrum were sorted. Negatively stained cells (P2, left-hand side 5%) were defined as CD133- and positively stained cells (P3, right-hand side 5%) were defined as CD133+ cells (A). CSPC marker genes were examined using quantitative real-time PCR, and the relative amount of each CSPC marker was calculated as 2^-ΔΔCt. The expression of Nanog, OCT-4, CD117, CD44, CD24, and CD133 in CD133– cells was compared with that of each gene in CD133+ cells. (B). MiR-21 expressed in CD133+ and CD133– cells. Quantitative real-time PCR was performed to detect expression of miR21. Expression of miR-21 in CD133– cells (basal level) was compared with the level of expression of miR-21 in CD133+ cells (C). The data in (B) and (C) represent the mean of three individual sets of experiments. The variations were from the calculation of standard deviation, and * indicates statistical significance at P-values less than 0.05. APC, allophycocyanin; CSPCs, cancer stem/progenitor cells; FACS, fluorescence-activated cell sorting; IgG, immunoglobulin G; MiR, micro RNA.