Fig. 2

KLF10 modulated stem cell phenotypes of PDAC. A Representative pictures of sphere formation (left panel) of Panc-1-pLKO and Pan-1-pLKO-shKLF10 cells. Original magnification: (left upper panels) 1 × 100, (left lower panels) 1 × 400. Quantitative bar plots (right panel) represent mean ± SE of cumulated data from three independent experiments. **p < 0.01. B Representative flow cytometry (left panel) of stem cell markers including CD44, CD24, c-MET and CD326 of Panc-1-pLKO and Panc-1-pLKO-shKLF10. Quantitative bar plots (right panel) represent mean ± SE of cumulated data from three independent experiments of flow cytometry measuring each stem cell markers. *p < 0.05 and **p < 0.01, respectively. C (Upper panel) immunoblots of KLF10 expression in MiaPaCa cells with conditional overexpression of KLF10 (pLVX-KLF10) as described in “Materials and methods”. Dox represents doxycylin. β-Actin was used as internal control. (Lower panel) Quantitative bar plots of sphere formation from three independent data of MiaPaCa cells with pLVX vector control or MiaPaCa-pLVX-KLF10 treated without or with Dox. **p < 0.01. D (Left upper panels) immunofluorescence staining of CD47 (green) in MiaPaCa-pLVX or MiaPaCa-pLVX-KLF10 without or with Dox. Nuclei were counterstained with DAPI. Quantitative bar plots (right upper panel) represent mean ± SE from three independent experiments. *p < 0.05 and **p < 0.01, respectively. (Lower left panels) representative flow cytometry of CD24 expression of MiaPaCa-pLVX, and MiaPaCa-pLVX-KLF10 without or with Dox treatment. (Lower right panel) quantitative bar plots represent mean ± SE from three independent experiments. **p < 0.01. E Representative immunoblots of KLF10 and phospho-AMPK in Panc-1 cells treated without or with 2 mM metformin and 0–30 µM Compound C as described in “Materials and methods”. F Representative images, ×100 (left upper panels) and ×400 (left lower panel), of sphere formation from Panc-1-pLKO and Panc-1-pLKO-shKLF10 treated without or with 2 mM metformin as described in “Materials and methods”. Quantitative bar plots (right panel) of mean ± SE from cumulated data of three independent experiments. *p < 0.05. G In vivo limiting dilution assay. Number of mice with tumor growth after implanted subcutaneously with Panc-1-pLKO and Panc-1-LKO-shKLF10 cells of various cell number indicated as described in “Materials and methods”. H Tumor growth curves of mice after 1 × 10⁴ cells of Panc-1-pLKO and Panc-1-pLKO-shKLF10 implanted. Each point represents mean ± SE of cumulated data from at least six mice. *p < 0.05