Journal:

Article Title: Adiponectin Modulates C-Jun N-Terminal Kinase and Mammalian Target of Rapamycin and inhibits hepatocellular carcinoma

doi: 10.1053/j.gastro.2010.07.001

Figure Lengend Snippet: A, Cells were serum starved for 16h followed by adiponectin treatment (0.5–10 μg/ml) for 24h and 48h and BrdU incorporation assays were performed. Adiponectin treatment decreased proliferation of HepG2 and Huh7 cells in a dose dependent manner whereas normal hepatocytes (THLE-2 and HH) remained unchanged. *p< 0.01, for different dose compared with untreated cells. The data represents mean values ± SEM and are the results of three independent experiments performed in triplicates. B and C, Cells were serum starved for 16h and treated with adiponectin (10μg/ml) (A) for 12h. Untreated cells (U), cells grown in complete medium containing 10% FBS (S) and cells treated with cycloheximide (C) were included as controls. Cycloheximide was employed as an inhibitor of proliferation. Total protein lysates were analyzed by immunoblot analysis with antibodies for cyclinD1 (B) and PCNA (C). Adiponectin treatment decreased expression of cyclinD1 and PCNA in HCC cells. The western blot signals were quantified by densitometry and normalized to β-actin. These data are representative of multiple independent experiments, *p< 0.05, compared with untreated controls.

Article Snippet: THLE-2 cells (ATCC) derived from primary normal liver cells and human hepatocytes (Lonza, Walkersville, MD) were used as controls.

Techniques: BrdU Incorporation Assay, Western Blot, Expressing

Journal:

Article Title: Adiponectin Modulates C-Jun N-Terminal Kinase and Mammalian Target of Rapamycin and inhibits hepatocellular carcinoma

doi: 10.1053/j.gastro.2010.07.001

Figure Lengend Snippet: A, Cells were serum-starved for 16h followed by adiponectin treatment (0.5–10 μg/ml) for indicated time-intervals and apoptosis was analyzed by XTT method. The percentage of apoptotic cells represents the means ± SEM of viable cells present in treatment calculated with respect to cells grown in complete medium. *p< 0.01 as compared to respective untreated cells. Adiponectin induced apoptosis in HepG2 and Huh7 cells in a dose and time dependent manner, while no apoptosis was observed in THLE-2 and human hepatocytes. B, HepG2 and Huh7 cells were serum-starved for 16h followed by adiponectin (10μg/ml) treatment (A) for 12 hours and caspase-3 like activity was analyzed. Untreated cells (U), cells treated with cycloheximide (C) and cells treated with caspase-3 inhibitor Z-VAD-FMK (carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]- fluoromethylketone) (Z) were included as controls. Adiponectin treatment increased caspase-3 like activity in HCC cells. The numbers in the figure represent the mean value of three repeat samples. C, Cells were treated as described in B. Cycloheximide was employed as an inhibitor of proliferation. Total protein lysates were analyzed by immunoblot analysis using caspase-3 and cleaved caspase-3 antibodies. The western blot signals were quantified by densitometry and normalized to β-actin. These data are representative of multiple independent experiments, *p < 0.05, compared with untreated controls.

Article Snippet: THLE-2 cells (ATCC) derived from primary normal liver cells and human hepatocytes (Lonza, Walkersville, MD) were used as controls.

Techniques: Activity Assay, Western Blot