(A–B), Effects of SFN on expression of Hh target genes in the pancreatic CSCs. Real time PCR (q-RT-PCR) was performed to examine the expression of Nanog and Oct4 and data were normalized with GAPDH. All assays were performed in triplicate and were calculated on the basis of ΔΔCt method. Data represent mean ± SD. @ and % = significantly different from control, P < 0.05. (C), Pancreatic CSCs were treated with SFN (0–20 µM), and cell lysates were collected and Western blot analysis was performed using anti- Nanog, Oct4 or β-actin antibody. (D–E), Effects of SFN on expression of Hh target genes involved in cell proliferation in the pancreatic CSCs. Real time PCR (q-RT-PCR) was performed to examine the expression of PDGFRα and Cyclin D1, involved in the maintenance of proliferation was analyzed and normalized with GAPDH. All assays were performed in triplicate and were calculated on the basis of ΔΔCt method. Data represent mean ± SD. @, %, and $ = significantly different from control, P < 0.05. (F), Pancreatic CSCs were treated with SFN (0–20 µM), and cell lysates were collected and Immunobloted for anti- PDGFRα, Cyclin D1 or β-actin antibody.