Journal: Biomolecules & Therapeutics

Article Title: ATF3 Mediates Anti-Cancer Activity of Trans -10, cis -12-Conjugated Linoleic Acid in Human Colon Cancer Cells

doi: 10.4062/biomolther.2014.107

Figure Lengend Snippet: t 10, c 12-CLA increases ATF3 expression and apoptosis in different human colorectal cancer cells. (A, C) HCT116, LoVo and HT-29 cells were incubated with media containing indicated concentrations of t 10, c 12-CLA for 24 h. Western analysis was performed for ATF3, cleaved PARP, caspase-3, and actin as described in Methods . Data represent one experiment. (B) Apoptosis of LoVo and HT-29 cells were analyzed with Cell Death Detection ELISA PLUS Kit (Roche Diagnostics). * p <0.05 versus vehicle-treated cells.

Article Snippet: HCT-116 cells were transfected with control or ATF3 small interference RNA (siRNA; Santa Cruz; Cat# sc-29757) at a concentration of 100 nM using TransIT-TKO transfection reagent (Mirus, Madison, WI, USA), as described previously ( Yamaguchi et al ., 2004 ).

Techniques: Expressing, Incubation, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Biomolecules & Therapeutics

Article Title: ATF3 Mediates Anti-Cancer Activity of Trans -10, cis -12-Conjugated Linoleic Acid in Human Colon Cancer Cells

doi: 10.4062/biomolther.2014.107

Figure Lengend Snippet: t 10, c 12-CLA increases transcriptional activity of ATF3 gene. (A) HCT-116 cells were treated with different doses of t 10, c 12-CLA for 24 h. Total RNA was isolated and RT-PCR was performed as described in Methods . Data represent one experiment. (B) The pATF3-1850/+34 construct (0.5 μg) was co-transfected with pRL-null vector (0.05 μg). The cells were treated with indicated doses of t 10, c 12-CLA for 24 h and then luciferase activity was measured as described in Methods . Values are expressed as mean ± SD of 3 replicates. * p <0.05 versus vehicle-treated cells. (C) HCT116 cells were transfected with indicated ATF3 deletion promoter constructs (0.5 μg) with pRL-null vector (0.05 μg). The cells were treated with vehicle or 50 μM of t 10, c 12-CLA for 24 h and luciferase activity was measured. Values are expressed as mean ± SD of 3 replicates. * p <0.05 versus vehicle-treated cells.

Article Snippet: HCT-116 cells were transfected with control or ATF3 small interference RNA (siRNA; Santa Cruz; Cat# sc-29757) at a concentration of 100 nM using TransIT-TKO transfection reagent (Mirus, Madison, WI, USA), as described previously ( Yamaguchi et al ., 2004 ).

Techniques: Activity Assay, Isolation, Reverse Transcription Polymerase Chain Reaction, Construct, Transfection, Plasmid Preparation, Luciferase

Journal: Biomolecules & Therapeutics

Article Title: ATF3 Mediates Anti-Cancer Activity of Trans -10, cis -12-Conjugated Linoleic Acid in Human Colon Cancer Cells

doi: 10.4062/biomolther.2014.107

Figure Lengend Snippet: t 10, c 12-CLA does not influence ATF3 stability (A) HCT-116 cells were treated with vehicle or t 10, c 12-CLA for 24 h, and subsequently co-treated with actinomycin D. At the indicated times, total RNAs were isolated and mRNA was examined by RT-PCR. (B) The band signals were quantified with Scion Image software. The relative level of ATF3 mRNA (relative to the level of GAPDH) was calculated and the results plotted as the % of the mRNA level present at time 0 of actinomycin D treatment. Data represent one experiment.

Article Snippet: HCT-116 cells were transfected with control or ATF3 small interference RNA (siRNA; Santa Cruz; Cat# sc-29757) at a concentration of 100 nM using TransIT-TKO transfection reagent (Mirus, Madison, WI, USA), as described previously ( Yamaguchi et al ., 2004 ).

Techniques: Isolation, Reverse Transcription Polymerase Chain Reaction, Software

Journal: Biomolecules & Therapeutics

Article Title: ATF3 Mediates Anti-Cancer Activity of Trans -10, cis -12-Conjugated Linoleic Acid in Human Colon Cancer Cells

doi: 10.4062/biomolther.2014.107

Figure Lengend Snippet: Dependency of ATF3 in t 10, c 12-CLA-induced GSK3β and NAG-1 expression, and apoptosis (A) HCT116 cells were exposed to different doses of t 10, c 12-CLA for 24 h and Western blot was performed GSK3β and actin. (B) The HCT116 (upper) and HT-29 (lower) cells were treated with vehicle or t 10, c 12-CLA (50 μM) for 24 h and stimulated with IGF-1 (100 ng/mL). Western analysis was performed using antibodies for phospho-Akt (Ser 473 ), total Akt and actin. (C, D) HCT-116 cells were transfected with control or ATF3 siRNA (100 nM) for 24 h and treated with t 10, c 12-CLA (50 μM) for 24 h. Western analysis was performed for ATF3, NAG-1, GSK3β, PARP, and actin. Data represent one experiment.

Article Snippet: HCT-116 cells were transfected with control or ATF3 small interference RNA (siRNA; Santa Cruz; Cat# sc-29757) at a concentration of 100 nM using TransIT-TKO transfection reagent (Mirus, Madison, WI, USA), as described previously ( Yamaguchi et al ., 2004 ).

Techniques: Expressing, Western Blot, Transfection, Control