Journal: PLoS ONE

Article Title: Resveratrol Mediated Modulation of Sirt-1/Runx2 Promotes Osteogenic Differentiation of Mesenchymal Stem Cells: Potential Role of Runx2 Deacetylation

doi: 10.1371/journal.pone.0035712

Figure Lengend Snippet: Whole cell lysates (500 ng of protein per lane) were probed with antibodies for collagen type I (a), for the osteogenic specific transcription factor Runx2 (b) and for the adipogenic specific transcription factor PPAR-γ (c) in MSC (A) and in pre-osteoblastic cells in high-density culture (B). Cultures were treated with 0.1, 1 and 10 µM resveratrol alone, or with 1, 10 and 100 mM nicotinamide alone or pre-treated with 1 µM resveratrol for 4 hours and then co-treated with 1, 10, 100 mM nicotinamide or left untreated for 2 weeks with osteogenic induction medium in high-density cultures. Untreated cultures (without resveratrol or nicotinamide) produced collagen type I (a, A–B) and Runx2 (b, A–B) in both cultures. Incubation with nicotinamide reduced collagen type I and Runx2 production and increased the expression of PPAR-γ in a concentration dependent manner in MSC-cultures (c, A) and decreased the expression of PPAR-γ in a concentration dependent manner in pre-osteoblastic cultures (III, B). However, pre-treatment with resveratrol inhibited the adverse effects of nicotinamide and the osteoblasts produced large amounts of collagen type I and Runx2. Synthesis of the housekeeping protein β-actin was unaffected (d, A–B).

Article Snippet: Polyclonal anti-PPAR-γ antibodies were purchased from Acris Antibodies GmbH, Germany.

Techniques: Produced, Incubation, Expressing, Concentration Assay

Journal: PLoS ONE

Article Title: Resveratrol Mediated Modulation of Sirt-1/Runx2 Promotes Osteogenic Differentiation of Mesenchymal Stem Cells: Potential Role of Runx2 Deacetylation

doi: 10.1371/journal.pone.0035712

Figure Lengend Snippet: Cultures were treated with 0, 1, 10, 100 mM nicotinamide or pre-treated with 1 µM resveratrol for 4 h followed by co-treatment with nicotinamide over 14 days with osteogenic induction medium. Cultures were lysed and immunoprecipitated with anti-PPAR-γ (a), or anti-Sirt-1 (b, c). The immunoprecipitates were separated by SDS-PAGE and analyzed by immunoblotting using anti-NCoR (a, b) and anti- PPAR-γ (c). The same blots were re-probed with an antibody to anti-PPAR-γ (a), anti-Sirt-1 (b, c). Results shown are representative of three independent experiments.

Article Snippet: Polyclonal anti-PPAR-γ antibodies were purchased from Acris Antibodies GmbH, Germany.

Techniques: Immunoprecipitation, SDS Page, Western Blot

Journal: PLoS ONE

Article Title: Resveratrol Mediated Modulation of Sirt-1/Runx2 Promotes Osteogenic Differentiation of Mesenchymal Stem Cells: Potential Role of Runx2 Deacetylation

doi: 10.1371/journal.pone.0035712

Figure Lengend Snippet: Cells were either untreated or treated with resveratrol (1 µM), nicotinamide (10 mM) or with Sirt-1 antisense (1 µM) or sense oligonucleotides (1 µM) in the presence of lipofectin alone or cells were pre-treated with resveratrol for 4 h followed by co-treatment with Sirt-1 antisense or sense oligonucleotides in the presence of lipofectin for 24 h or/and with nicotinamide over 21 days with osteogenic induction medium in monolayer cultures. (A) Whole cell lysates (500 ng/lane) were fractionated and subjected to western blotting with antibodies against Sirt-1 and β-actin. Synthesis of the housekeeping protein β-actin was unaffected. (B) Whole-cell extracts were prepared, immunoprecipitated with an anti-Runx2 antibody, and subjected to western blot analysis using an anti–acetyl-lysine antibody. The same blots were re-probed with an antibody to anti-Runx2. Whole cell lysates (500 ng/lane) were fractionated and analyzed by immunoblotting using anti-osteocalcin (C) or anti-PPAR-γ (D) antibodies and β-actin. Synthesis of the housekeeping protein β-actin was unaffected.

Article Snippet: Polyclonal anti-PPAR-γ antibodies were purchased from Acris Antibodies GmbH, Germany.

Techniques: Western Blot, Immunoprecipitation

Journal: Experimental and Therapeutic Medicine

Article Title: Effects of curcumin on the apoptosis of cardiomyocytes and the expression of NF-κB, PPAR-γ and Bcl-2 in rats with myocardial infarction injury

doi: 10.3892/etm.2016.3858

Figure Lengend Snippet: Optical density of NF-κB and PPAR-γ protein expression in the cardiomyocytes of rats (mean ± standard deviation).

Article Snippet: Following blocking with 10% goat serum at 37°C for 15 min, the slices were incubated with rabbit anti-PPAR-γ polyclonal antibody (Peprotech Inc., Rocky Hill, NJ, USA; 1:200) or rabbit anti-NF-κBp6 polyclonal antibody (Peprotech Inc.; 1:100) at 4°C overnight.

Techniques: Expressing, Control