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. 2014 Nov 26;6(2):651–661. doi: 10.18632/oncotarget.2892

Figure 3. Gene silencing of GLUT1 by siRNA causes loss of the self-renewal capacity of CSCs.

Figure 3

(A) Cells were transiently transfected with an siRNA against GLUT1 (siGLUT1-#3), and as controls, with an siRNA against Lamin A/C (siLamin A/C) or with a non-targeting siRNA (siControl), as detailed in Materials and methods. The transfected cells were then subjected, 6 days after transfection for PANC-1 CSLCs and 8 days after transfection for A2780 CSCs and GS-Y03 cells, to immunoblot analyses for the expression of the indicated proteins. (B) Cells treated as in (A) were subjected to flow cytometric analysis for the cell surface expression of CD133. Representative flow cytometric plots together with the percentages of CD133-positive cells are shown. (C) Cells treated as in (A) were subjected to the sphere formation assay. Left panels show the photographs of representative wells. The graphs show the percentage of wells in which a tumor sphere was formed from a single cell. Values represent means + SD from three independent experiments. *p<0.01. Bar: 50 μm.