Journal: International Journal of Molecular Sciences

Article Title: Effects of Maternal Resveratrol on Maternal High-Fat Diet/Obesity with or without Postnatal High-Fat Diet

doi: 10.3390/ijms21103428

Figure Lengend Snippet: Primary and secondary antibodies used for Western blot.

Article Snippet: pPPARγ (phosphoryl peroxisome proliferator-activated receptors γ) , Bioss , bs-3737R , 1:1000.

Techniques: Western Blot

Journal: International Journal of Molecular Medicine

Article Title: Adropin attenuates pancreatitis-associated lung injury through PPARγ phosphorylation-related macrophage polarization

doi: 10.3892/ijmm.2023.5298

Figure Lengend Snippet: Excessive M1 macrophage polarization is observed in the Adro-KO + L-Arg group. (A) Western blot analysis of lung tissue form the Adro-KO + L-Arg group. (B) Co-expression of CD68 and iNOS in lung tissue in L-Arg group determined using immunofluorescence. (C) Co-expression of CD68 and CD206 of lung tissue in the L-Arg group determined using immunofluorescence. (D) iNOS mRNA level in lung tissue from the Adro-KO + L-Arg group (n≥8). (E) CD68 mRNA level in lung tissue from the Adro-KO + L-Arg group (n≥7). (F) CD163 mRNA level in lung tissue from the Adro-KO + L-Arg group (n≥8). (G) Arg-1 mRNA level in lung tissue from the Adro-KO + L-Arg group (n≥8). (H) Quantitative analysis of immunofluorescence (CD68 + iNOS) staining (n≥5). (I) Quantitative analysis of immunofluorescence (CD68; CD206) staining in macrophages (n≥9). (J) Quantitative analysis of the western blots (PPARγ; n≥6); (K) Quantitative analysis of the western blots (PPARγ Ser273; n≥6). (L) Quantitative analysis of the western blots (PPARγ Ser112; n≥6). * P<0.05. L-Arg, L-arginine; NS, normal saline; Adro-KO, adropin knockout; iNOS, inducible nitric oxide synthase.

Article Snippet: The primary antibodies were used at 4°C overnight, and the antibody included Adropin (1:1,000, cat. no. PA5-72781, Thermo Fisher Scientific, Inc.); GADPH (1:10,000; cat. no. AC001, ABclonal); peroxisome proliferator-activated receptor γ (PPARγ; 1:1,000, cat. no. bsm-52220R, BIOSS); p-PPARγ Ser112 (1:1,000, cat. no. bs-3737R, BIOSS); p-PPARγ Ser273 (1:1,000, cat. no. bs-2875R, BIOSS), caspase-3 (1:1,000, cat. no. YC0006, ImmunoWay Biotechnology Company) and PARP1 (1:1,000, cat. no. A0942, ABclonal).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Saline, Knock-Out

Journal: International Journal of Molecular Medicine

Article Title: Adropin attenuates pancreatitis-associated lung injury through PPARγ phosphorylation-related macrophage polarization

doi: 10.3892/ijmm.2023.5298

Figure Lengend Snippet: Adropin exogenous supplement induces M2 macrophage polarization. (A) Western blot analysis of lung tissue from the Adro-KO + L-Arg + Adro (34−76) group. (B) Quantitative analysis of the western blots (PPARγ, PPARγ Ser112, PPARγ Ser273) (n≥5). (C) Co-expression of CD68 and iNOS in lung tissue from the Adro-KO + L-Arg+Adro (34−76) group. (D) Co-expression of CD68 and iNOS in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group. (E) Ratio of CD68/DAPI of lung in Adro-KO+ L-Arg+Adro (34−76) group(n≥3); (F) The ratio of iNOS/DAPI in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥3). (G) Ratio of iNOS/CD68 in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥3). (H) Ratio of CD68/DAPI in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥3). (I) Ratio of CD206/DAPI in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥3). (J) Ratio of CD206/CD68 in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥3). (K) CD163 mRNA level in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥6). (L) Arg-1 mRNA level in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥6). (M) iNOS mRNA level in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥6). (N) CD86 mRNA level level in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥6). * P<0.05 and *** P<0.001. L-Arg, L-arginine; Adro-KO, adropin knockout; NS, normal saline; iNOS, inducible nitric oxide synthase; Arg-1, arginase 1.

Article Snippet: The primary antibodies were used at 4°C overnight, and the antibody included Adropin (1:1,000, cat. no. PA5-72781, Thermo Fisher Scientific, Inc.); GADPH (1:10,000; cat. no. AC001, ABclonal); peroxisome proliferator-activated receptor γ (PPARγ; 1:1,000, cat. no. bsm-52220R, BIOSS); p-PPARγ Ser112 (1:1,000, cat. no. bs-3737R, BIOSS); p-PPARγ Ser273 (1:1,000, cat. no. bs-2875R, BIOSS), caspase-3 (1:1,000, cat. no. YC0006, ImmunoWay Biotechnology Company) and PARP1 (1:1,000, cat. no. A0942, ABclonal).

Techniques: Western Blot, Expressing, Knock-Out, Saline

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