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. Author manuscript; available in PMC: 2014 Oct 1.

Published in final edited form as: J Cell Physiol. 2013 Oct;228(10):1996–2005. doi: 10.1002/jcp.24362

(A) and (B) HuH7 cells were treated with sorafenib (Sor, 3 µM) and/or Na valproate (Val, 1 mM). In (A) cells were treated with increasing doses of TRAIL (0–5 ng/ml). Twelve h after treatment cells were isolated and the levels of cytochrome c released into the cytosolic fraction determined. In (B) cells were treated with increasing doses of obatoclax (GX, 0–50 nM). Twelve h after treatment cells were isolated and the levels of cytochrome c released into the cytosolic fraction determined. The –Fold increase in cytosolic cytochrome c levels were determined (n = 3 +/− SEM). **p < 0.05 greater fold increase release than in GX treated cells. Errors are not shown due to space restrictions.(C) HEP3B cells were transfected with scrambled siRNA (siSCR) or siRNA molecules to knock down BID, BAX or NOXA expression. Twenty four h after treatment cells were treated with vehicle (DMSO+PBS) or sorafenib (Sor, 3 µM) plus valproate (1 mM) plus TRAIL (5 ng/ml). Cells were isolated 24h after drug exposure and cell viability determined by trypan blue exclusion (n = 3 +/− SEM).#p < 0.05 value less than in siSCR cells. (D) HEP3B cells were transfected with scrambled siRNA (siSCR) or siRNA molecules to knock down BID, BAX or NOXA expression. Twenty four h after treatment cells were treated with vehicle (DMSO+PBS) or sorafenib (Sor, 3 µM) plus valproate (1 mM) plus obatoclax (50 nM). Cells were isolated 24h after drug exposure and cell viability determined by trypan blue exclusion (n = 3 +/− SEM). #p < 0.05 value less than in siSCR cells. (E) HEPG2; (F) HEP3B; (G) HuH7 cells were infected with empty vector virus Ad.cmv or viruses to express c-FLIP-s, BCL-XL or dominant negative caspase 9. Twenty four h after infection cells were treated with vehicle (DMSO) or sorafenib (Sor, 3 µM) and Vorinostat (Vor, 500 nM)and/or TRAIL (5 ng/ml). Cells were isolated 24h after drug exposure and cell viability determined by trypan blue exclusion (n = 3 +/− SEM). #p < 0.05 value less than in CMV cells. (H) Hepatoma cells were transfected with empty vector plasmid (CMV) or a plasmid to express MCL-1.Twenty four h after transfection cells were treated with vehicle (DMSO) or sorafenib (Sor, 3 µM) and Vorinostat (Vor, 500 nM)and TRAIL (5 ng/ml). Cells were isolated 24h after drug exposure and cell viability determined by trypan blue exclusion (n = 3 +/− SEM). #p < 0.05 value less than in CMV cells.

(A) and (B) HuH7 cells were treated with sorafenib (Sor, 3 µM) and/or Na valproate (Val, 1 mM). In (A) cells were treated with increasing doses of TRAIL (0–5 ng/ml). Twelve h after treatment cells were isolated and the levels of cytochrome c released into the cytosolic fraction determined. In (B) cells were treated with increasing doses of obatoclax (GX, 0–50 nM). Twelve h after treatment cells were isolated and the levels of cytochrome c released into the cytosolic fraction determined. The –Fold increase in cytosolic cytochrome c levels were determined (n = 3 +/− SEM). **p < 0.05 greater fold increase release than in GX treated cells. Errors are not shown due to space restrictions.(C) HEP3B cells were transfected with scrambled siRNA (siSCR) or siRNA molecules to knock down BID, BAX or NOXA expression. Twenty four h after treatment cells were treated with vehicle (DMSO+PBS) or sorafenib (Sor, 3 µM) plus valproate (1 mM) plus TRAIL (5 ng/ml). Cells were isolated 24h after drug exposure and cell viability determined by trypan blue exclusion (n = 3 +/− SEM).#p < 0.05 value less than in siSCR cells. (D) HEP3B cells were transfected with scrambled siRNA (siSCR) or siRNA molecules to knock down BID, BAX or NOXA expression. Twenty four h after treatment cells were treated with vehicle (DMSO+PBS) or sorafenib (Sor, 3 µM) plus valproate (1 mM) plus obatoclax (50 nM). Cells were isolated 24h after drug exposure and cell viability determined by trypan blue exclusion (n = 3 +/− SEM). #p < 0.05 value less than in siSCR cells. (E) HEPG2; (F) HEP3B; (G) HuH7 cells were infected with empty vector virus Ad.cmv or viruses to express c-FLIP-s, BCL-XL or dominant negative caspase 9. Twenty four h after infection cells were treated with vehicle (DMSO) or sorafenib (Sor, 3 µM) and Vorinostat (Vor, 500 nM)and/or TRAIL (5 ng/ml). Cells were isolated 24h after drug exposure and cell viability determined by trypan blue exclusion (n = 3 +/− SEM). #p < 0.05 value less than in CMV cells. (H) Hepatoma cells were transfected with empty vector plasmid (CMV) or a plasmid to express MCL-1.Twenty four h after transfection cells were treated with vehicle (DMSO) or sorafenib (Sor, 3 µM) and Vorinostat (Vor, 500 nM)and TRAIL (5 ng/ml). Cells were isolated 24h after drug exposure and cell viability determined by trypan blue exclusion (n = 3 +/− SEM). #p < 0.05 value less than in CMV cells.