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. 2021 Jun 23;13(13):3145. doi: 10.3390/cancers13133145

Figure 2.

Figure 2

Interplay between pluripotency/stemness factors and telomerase activity in pancreatic CSCs: (A,B) RT-qPCR analysis of pluripotency/stemness-associated genes in primary pancreatic CSCs enriched by CD133 expression (A) or ALDEFLUOR activity (B) vs. CD133− and ALDEFLUOR- non-CSCs (n = at least 3 independent FACSortings). (C–F) Quantitative RT-PCR (C), telomerase activity (D), flow cytometry analysis for CD133 (E) and sphere formation capability (F), compared in NANOG negative cells (white) vs. NANOG positive cells (yellow) using a NANOG-YNL reporter. (G) Schematic illustration and quantification on the production of virions in parental pseudorabies viruses (PRV-NIA3 and vBecker) compared to telomerase activity-dependent virus production (PRV-TER) in adherent HPDE and Panc185 cells as compared to Panc185 spheres. (H) Clonogenic Assay in NANOG-YNL positive and negative cells after withdrawing BIBR1532 (n = ≥3 independent experiments. ns = not statistically significant) (I) Gene expression levels of stemness/pluripotency genes and TERT in HEK293T cells after inducing expression of SOX2, OCT3/4, KLF4 and NANOG, compared to the empty vector control (n = 4 independent experiments). In (A–I), data are represented as mean ± SEM. * p ≤ 0.05 (Mann–Whitney-U test). (J) Telomerase activity after inducing expression of SOX2, OCT3/4, KLF4 and NANOG, compared to the empty vector control. Representative TRAP assays and quantification are depicted.