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. 2009 Dec 9;285(7):4931–4940. doi: 10.1074/jbc.M109.048397

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FIGURE 3. — SP cells induction with stable transfection of Mad2 in human NPC cells. A, Hoechst 33342 staining was performed in human NPC CNE-2-S22-Mad2 cells. a, parental CNE-2-S22 cells; b, Mad2-transfected CNE-2-S22 cells; c, CNE-2-S22-Mad2 cells transfected with scrambled siRNA; d, CNE-2-S22-Mad2 cells transfected with siMad2. e–h, the same as a–d, except that FTC at a final concentration of 5 μm was added before Hoechst 33342 staining. The percentage of SP cells is indicated. B, spheroid (nasosphere) formation assay of CNE-2-S22 and CNE-2-S22-Mad2 in suspension culture circumstance. C, colony formation assay of CNE-2-S22 and CNE-2-S22-Mad2 in adherent culture circumstance. The p value was indicated in the figures. D, Western blot analysis of exogenous Mad2 tagged with Myc and endogenous Sox2 in CNE-2-S22 and CNE-2-S22-Mad2 stable cell lines. E, Western blot analysis of endogenous Mad2. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control.