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. 2012 Sep 28;7(9):e46083. doi: 10.1371/journal.pone.0046083

Figure 5. Regulation of Gli on spheroid formation by SFN in pancreatic CSCs in vitro.

Figure 5

(A), Inhibitory effects of SFN on CSC's spheroid viability. Pancreatic CSCs were seeded as described above and treated with SFN (0 and 20 µM). After 7 and 14 days, primary and secondary spheroids were dissociated and viable cells were counted by trypan blue assay. Data represent mean ± SD. @, and $ = significantly different from control, P < 0.05. (B), Pancreatic CSCs were transduced with either scrambled shRNA or Gli1 and Gli2 shRNA expressing lentiviral vectors (pLKO.1), and cell lysates were collected and Western blot analysis was performed using anti-Gli1 or Gli2 antibody. (C), CSC/scrambled and CSC/Gli shRNA were seeded as described above and treated with SFN (0–20 µM). After 7 days, spheroids were collected and cell suspensions were prepared and viable cells were counted by trypan blue assay. Data represent mean ± SD. @, %, and * = significantly different from control, P < 0.05. (D), Gli shRNA enhances the inhibitory effects of SFN on CSC's spheroid viability. CSC/scrambled and CSC/Gli shRNA were seeded as described above and treated with SFN (0 and 20 µM). After 7 days, spheroids were photomicrographed.