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Fig. 5 | Cancer Cell International

Fig. 5

From: R-spodin2 enhances canonical Wnt signaling to maintain the stemness of glioblastoma cells

Fig. 5

Wnthigh and Wntlow sub-populations of U251 cells exhibit different cellular properties in vitro. a Isolation of Wnthigh and Wntlow sub-populations of U251 cells. Left panel, Typical flow cytometry analysis depicting unsorted U251-7TGP cells treated with 20 ng/ml Wnt3A (yellow). For subsequent experiment, the 5% of cells with the highest and lowest GFP levels were sorted. Right panel, shown is the flow cytometric reanalysis of the respective subgroups after several rounds of enrichment. b Real-time PCR demonstrates that Wnthigh cells show higher expression levels of β-catenin targets and stem cell markers. c Sphere formation assay shows that Wnthigh cells have an enhanced sphere formation ability. Scale bar is 200 μm. Quantification analysis of data is expressed as the Mean ± SD from three independent experiments. d Soft agar assay shows Wnthigh cells have a higher anchorage-independent cell growth ability. Quantification analysis of data is expressed as the Mean ± SD from three independent experiments. e Transwell assay shows Wnthigh cells exhibit enhanced cell migratory ability. Quantification analysis of data is expressed as the Mean ± SD from three independent experiments. N. C. indicates vehicle control of Wnthigh and Wntlow cells

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