Figure 2

EcPDF expression induces CSC-like phenotypes in colorectal cancer cells. (A) Morphologies of control and EcPDF_3f SW480 cells. F-actin and nuclei were stained using Alexa Fluor 594-conjugated phalloidin and DAPI, respectively. Scale bars, 50 μm. (B) Quantification of cell sizes in (A). (C) Spheroid analyses of control and EcPDF_3f SW480 cells. Spheroids with a diameter exceeding 50 μm were counted. Scale bars, 100 μm. (D) Quantification of data in (C). (E) The levels of CD24 mRNA in control and EcPDF_3f SW480 cells. (F) A representative FACS histogram depicting non-specific (isotype) and specific CD24 fluorescence patterns in control and EcPDF_3f SW480 cells. (G) The levels of SOX2 mRNA in control and EcPDF_3f SW480 cells. (H) Immunoblotting of SOX2 in control and EcPDF_3f SW480 cells with or without actinonin (2 µg/ml) treatment for 2 days. (I) Quantification of data in (H). (J) The growth of vector only-expressing (control) and EcPDF_3f-expressing HT29 cells. (K) Immunoblotting of SOX2 in control and EcPDF_3f-expressing HT29 cells. (L) Same as in (F) but with control and EcPDF_3f-expressing HT29 cells. In (B,D,E,G,I,J), error bars are presented as the mean ± SD of three replicate experiments; two-tailed t-test; significant P values are indicated in the figures.