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. Author manuscript; available in PMC: 2007 Apr 1.
Published in final edited form as: Gastroenterology. 2006 Apr;130(4):1283–1300. doi: 10.1053/j.gastro.2006.01.010

Fig. 8. Diminished HNF6 levels in Hepa1-6 cells by siRNA transfection causes a significant decrease in DNA replication following serum stimulation.

Fig. 8

Mouse hepatoma Hepa1-6 cells were transfected with HNF6 siRNA, control siRNA or left untransfected, serum starved for 48 hours and then stimulated to reenter the cell cycle with the addition of 10% fetal calf serum (FCS). These siRNA transfected or untreated Hepa1-6 cells were subjected to a one-hour pulse label with BrdU prior to harvesting them at 16 hours after serum stimulation and then DNA replication was determined by immunostaining of cells with BrdU antibody. (A–B) RNase protection assays and Western blot analysis demonstrated that transfection of Hepa1-6 cells with HNF6 siRNA caused a 50% reduction in levels of HNF6 mRNA and protein, but HNF6 expression was not influenced by transfection with a control siRNA. (C) Graph depicting reduced number of BrdU positive Hepa1-6 cells depleted in HNF6 levels following serum stimulation. Diminished number of BrdU positive Hepa1-6 cells after transfection of HNF6 siRNA to diminish HNF6 expression compared to Hepa1-6 cells transfected with control siRNA or left untransfected. Quantitation of the BrdU incorporation rates by counting the number of BrdU positive Hepa1-6 cells in 5 fields from three different HNF6 siRNA transfected cell plates. Student T-test determines statistically significant decrease in BrdU incorporation. The asterisks indicate statistically significant changes: **P ≤ 0.01 and P ≤ 0.001.