Fig. 3.

Characterization of reprogrammed PDAC cells generated by transfection with episomal vectors. a Cells from PDAC-247 were reprogrammed and different passages of the iPS-like clones are shown. b ALP activity was only observed in a few of the screened colonies from 247- reprogrammed cells; scale bar: 50 μm. c Immunofluorescence staining of pluripotency markers NANOG, TRA-1-81, SOX2, OCT4 and TRA-1-60 in the 247-parental and reprogrammed cells (upper panel). Both parental and reprogrammed cells were negative for SOX2, OCT4 and TRA-1-60 (lower panel). DAPI was used for nuclear counterstaining; scale bar: 100 μm. d Expression of pluripotency markers and epigenetic modifier genes in parental and reprogrammed PDAC-247 cells as assessed by real-time PCR. Gene expression levels were normalized to bACTIN; *p ≤ 0.0001; mean ± SD; n = 3. e Detection of NANOG mRNA expression using SmartFlare mRNA probe for NANOG in live parental and reprogrammed PDAC-247 cells. Inside a single colony, expression of NANOG is more pronounced in some areas. The circular binding pattern of the SmartFlare mRNA probe is typical for live imaging of NANOG; scale bar: 100 μm. f Expression of CD133 in parental and reprogrammed 247 cells by real-time PCR. Gene expression levels were normalized to bACTIN; *p ≤ 0.0001, mean ± SD; n = 3. g Representative flow cytometry images for CD133 staining