rabbit polyclonal antibody against pparγ  (Proteintech)

Journal: Nature Communications

Article Title: Quinolinate promotes macrophage-induced immune tolerance in glioblastoma through the NMDAR/PPARγ signaling axis

doi: 10.1038/s41467-023-37170-z

Figure Lengend Snippet: a Macrophages obtained from C57BL/6 mice were polarized towards the M2 phenotype ±QA (20 µM) or the PPARγ agonist troglitazone (Trog; 5 µM) and evaluated for the indicated proteins by western blot, which is representative of three biologically independent experiments. b Macrophages obtained from C57BL/6 mice were polarized towards the M2 phenotype ±Trog ( n = 5), GW9662 (PPARγ antagonist; n = 3) and/or QA (20 µM; n = 5) and analyzed for M2 macrophage markers (CD45 + CD11b + F4/80 + CD206 + ) by flow cytometry. All samples were biologically independent. Numbers represent mean ± SD. c PPARγ transcriptional activity was evaluated in the murine macrophage cell line IC-21 polarized towards the M2 phenotype ±siRNA of indicated proteins by PPARγ transcriptional activity ELISA ( n = 5 biologically independent samples). d C57BL/6 mouse-derived macrophages were polarized towards the M2 phenotype ±QA (20 µM) and evaluated for the indicated proteins by western blot, which is representative of 3 biologically independent experiments. e Macrophages were cultured in the presence or absence of QA ± the Foxo1 inhibitor AS-1842856 (0.1 µM) and evaluated for M2 macrophage markers (CD45 + CD11b + F4/80 + CD206 + ) by flow cytometry. Numbers represent mean ± SD ( n = 4 biologically independent samples/group). f Chromatin immunoprecipitation (ChIP) was performed on C57BL/6 mouse-derived macrophages polarized towards the M2 phenotype and evaluated alone (gray), with QA (red) or with Trog (blue). ChIP was performed using antibodies against PPARγ, Foxo1, and histone 3 (positive control), and IgG antibody (negative control). After IP and DNA purification, the PPARγ exon (202 bp in length) was analyzed using qPCR and gel electrophoresis. Box plots represent interquartile range, line between the data points represents mean, and whiskers represents SE ( n = 4 biologically independent samples/group). Statistics: One-way ANOVA followed by Tukey’s multiple comparisons test ( b , c ), two-tailed Student’s t -test ( e , f ). All tests were performed at 95% confidence interval. Source data are provided as a source data file.

Article Snippet: Chromatin was sonicated and then incubated with a polyclonal antibody against PPARγ (81B8 and C26H12) or FoxO1 (C29H4) Cell Signaling Technology (Danvers, MA) for immunoprecipitation.

Techniques: Western Blot, Flow Cytometry, Activity Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Cell Culture, Chromatin Immunoprecipitation, Positive Control, Negative Control, DNA Purification, Nucleic Acid Electrophoresis, Two Tailed Test

Journal: Nature Communications

Article Title: Quinolinate promotes macrophage-induced immune tolerance in glioblastoma through the NMDAR/PPARγ signaling axis

doi: 10.1038/s41467-023-37170-z

Figure Lengend Snippet: a M2 macrophages were polarized ±QA, gated for macrophage markers (CD45 + CD11b + F4/80 + ) and CD206 + ve with (green) or without (red) QA or CD206-ve with (orange) or without (dark green) QA, and evaluated for expression of the NMDA receptor 1 (NMDAR1; n = 2 biologically independent samples/group). b M2 macrophages were cultured in ±QA, L-AP5 (NMDAR1 inhibitor), or NMDA and cells were gated for M2 macrophages markers (CD45 + CD11b + F4/80 + CD206 + ). Numbers represent mean ± SD ( n = 3 biologically independent samples/group). c M2 macrophages were cultured in ±QA, NMDA, or Trog, and indicated proteins were evaluated by western blot, which is representative of three biologically independent experiments. d Schematic depicting the proposed mechanism of QA-induced M2 macrophage polarization. QA binds to the NMDA receptor of macrophages (1), leading to phosphorylation of Foxo1 (2). Phosphorylated Foxo1 is retained in the cytoplasm and destined for ubiquitination. Loss of nuclear Foxo1, a negative regulator of the PPARγ promoter, leads to increased PPARγ expression and transcriptional programs designed to promote macrophage polarization towards the M2 phenotype (3). Statistics: Two-tailed Student’s t test at 95% confidence interval ( b ). Source data are provided as a source data file.

Article Snippet: Chromatin was sonicated and then incubated with a polyclonal antibody against PPARγ (81B8 and C26H12) or FoxO1 (C29H4) Cell Signaling Technology (Danvers, MA) for immunoprecipitation.

Techniques: Expressing, Cell Culture, Western Blot, Phospho-proteomics, Ubiquitin Proteomics, Two Tailed Test

Journal: Frontiers in Nutrition

Article Title: Fenofibrate enhances lipid deposition via modulating PPARγ, SREBP-1c, and gut microbiota in ob/ob mice fed a high-fat diet

doi: 10.3389/fnut.2022.971581

Figure Lengend Snippet: The primers used for the polymerase chain reaction (PCR) reaction.

Article Snippet: Then, the membranes were blotted with the corresponding primary and secondary antibodies listed as follows: a mouse monoclonal antibody against SREBP-1c (2A4) (SANTA CRUZ, USA, sc-13551, 1:500), a rabbit polyclonal antibody against PPARγ (Proteintech, USA, 16643-1-AP, 1:1000), a mouse monoclonal antibody against PPARα (Abcam, USA, ab97609, 1:800), a mouse monoclonal antibody against β-actin (Abcam, USA, ab8226, 1:1000), a rabbit polyclonal antibody against LDLR (Abcam, USA, ab30532, 1:200), and secondary antibodies including a goat anti-rabbit IgG H&L (HRP) (Abcam, USA, ab6721, 1:3000) and a goat anti-mouse IgG H&L (HRP) (Abcam, USA, ab205719, 1:5000).

Techniques: Polymerase Chain Reaction

Journal: Frontiers in Nutrition

Article Title: Fenofibrate enhances lipid deposition via modulating PPARγ, SREBP-1c, and gut microbiota in ob/ob mice fed a high-fat diet

doi: 10.3389/fnut.2022.971581

Figure Lengend Snippet: Effect of fenofibrate on the expression of proteins involved in lipid metabolism in the plasma and liver of the ob/ob mice ( n = 6). (A) protein expression and densitometric quantification of plasma apolipoprotein (Apo) A-I; (B) protein expression and densitometric quantification of plasma Apo B; (C) protein expression and densitometric quantification of low-density lipoprotein receptor (LDLR) in the liver; (D) protein expression and densitometric quantification of peroxisome proliferator activated receptor (PPAR) α in the liver; (E) protein expression and densitometric quantification of PPARγ in the liver; (F) protein expression and densitometric quantification of sterol regulatory element binding protein (SREBP)-1c in the liver. Data were expressed as mean ± SD, and the statistical analysis was performed by Student- t -test. * means significantly different at p < 0.05 vs . Vehicle group; ** means significantly different at p < 0.01 vs . Vehicle group. Vehicle: mice were treated with 0.5 mL of water by gavage for 13 weeks; Fenofibrate: mice were treated with 0.5 mL of fenofibrate suspension liquid at the dose of 20 mg/kg/d by gavage for 13 weeks.

Article Snippet: Then, the membranes were blotted with the corresponding primary and secondary antibodies listed as follows: a mouse monoclonal antibody against SREBP-1c (2A4) (SANTA CRUZ, USA, sc-13551, 1:500), a rabbit polyclonal antibody against PPARγ (Proteintech, USA, 16643-1-AP, 1:1000), a mouse monoclonal antibody against PPARα (Abcam, USA, ab97609, 1:800), a mouse monoclonal antibody against β-actin (Abcam, USA, ab8226, 1:1000), a rabbit polyclonal antibody against LDLR (Abcam, USA, ab30532, 1:200), and secondary antibodies including a goat anti-rabbit IgG H&L (HRP) (Abcam, USA, ab6721, 1:3000) and a goat anti-mouse IgG H&L (HRP) (Abcam, USA, ab205719, 1:5000).

Techniques: Expressing, Clinical Proteomics, Binding Assay, Suspension

Journal: Frontiers in Nutrition

Article Title: Fenofibrate enhances lipid deposition via modulating PPARγ, SREBP-1c, and gut microbiota in ob/ob mice fed a high-fat diet

doi: 10.3389/fnut.2022.971581

Figure Lengend Snippet: Effect of fenofibrate on the expression of genes involved in lipid metabolism in the liver of the ob/ob mice. Gene relative expression of (A) peroxisome proliferator activated receptor (PPAR) α ( n = 12); (B) acyl-CoA oxidase (ACOX)-1 ( n = 12); (C) carnitine palmitoyltransferase (CPT)-1α ( n = 12); (D) CPT-2 ( n = 12); (E) PPARγ ( n = 7); (F) sterol regulatory element binding protein (SREBP)−1c ( n = 9); (G) stearoyl Coenzyme A desaturase-1 (SCD-1) ( n = 4); (H) fatty acid synthase (FAS) ( n = 4). The RT-qPCR experiment was repeated for one more time to confirm the gene expression trends of PPARα, ACOX-1, CPT-1α, CPT-2, and SREBP-1c. Data were expressed as mean ± SD, and the statistical analysis was performed by Student- t -test. * means significantly different at p < 0.05 vs . Vehicle group; ** means significantly different at p < 0.01 vs . Vehicle group. Vehicle: mice were treated with 0.5 mL of water by gavage for 13 weeks; Fenofibrate: mice were treated with 0.5 mL of fenofibrate suspension liquid at the dose of 20 mg/kg/d by gavage for 13 weeks.

Article Snippet: Then, the membranes were blotted with the corresponding primary and secondary antibodies listed as follows: a mouse monoclonal antibody against SREBP-1c (2A4) (SANTA CRUZ, USA, sc-13551, 1:500), a rabbit polyclonal antibody against PPARγ (Proteintech, USA, 16643-1-AP, 1:1000), a mouse monoclonal antibody against PPARα (Abcam, USA, ab97609, 1:800), a mouse monoclonal antibody against β-actin (Abcam, USA, ab8226, 1:1000), a rabbit polyclonal antibody against LDLR (Abcam, USA, ab30532, 1:200), and secondary antibodies including a goat anti-rabbit IgG H&L (HRP) (Abcam, USA, ab6721, 1:3000) and a goat anti-mouse IgG H&L (HRP) (Abcam, USA, ab205719, 1:5000).

Techniques: Expressing, Binding Assay, Quantitative RT-PCR, Gene Expression, Suspension

Journal: Frontiers in Nutrition

Article Title: Fenofibrate enhances lipid deposition via modulating PPARγ, SREBP-1c, and gut microbiota in ob/ob mice fed a high-fat diet

doi: 10.3389/fnut.2022.971581

Figure Lengend Snippet: Effect of fenofibrate on the fat index and expression of proteins involved in lipid metabolism in the white adipose of the ob/ob mice ( n = 6 unless otherwise specialized). (A) fat index was calculated according to the following formula: fat index = [(white adipose weight × 100%) ÷ body weight] ( n = 7 for the Vehicle group and n = 8 for the Fenofibrate group). Protein expression and densitometric quantification (B) peroxisome proliferator activated receptor (PPAR) α; (C) PPARγ; (D) sterol regulatory element binding protein (SREBP)-1c. Data were expressed as mean ± SD, and the statistical analysis was performed by Student- t -test. ** means significantly different at p < 0.01 vs . Vehicle group. Vehicle: mice were treated with 0.5 mL of water by gavage for 13 weeks; Fenofibrate: mice were treated with 0.5 mL of fenofibrate suspension liquid at the dose of 20 mg/kg/d by gavage for 13 weeks.

Article Snippet: Then, the membranes were blotted with the corresponding primary and secondary antibodies listed as follows: a mouse monoclonal antibody against SREBP-1c (2A4) (SANTA CRUZ, USA, sc-13551, 1:500), a rabbit polyclonal antibody against PPARγ (Proteintech, USA, 16643-1-AP, 1:1000), a mouse monoclonal antibody against PPARα (Abcam, USA, ab97609, 1:800), a mouse monoclonal antibody against β-actin (Abcam, USA, ab8226, 1:1000), a rabbit polyclonal antibody against LDLR (Abcam, USA, ab30532, 1:200), and secondary antibodies including a goat anti-rabbit IgG H&L (HRP) (Abcam, USA, ab6721, 1:3000) and a goat anti-mouse IgG H&L (HRP) (Abcam, USA, ab205719, 1:5000).

Techniques: Expressing, Binding Assay, Suspension

Journal: Oncology Letters

Article Title: Activation of PPARγ in bladder cancer via introduction of the long arm of human chromosome 9

doi: 10.3892/ol.2022.13212

Figure Lengend Snippet: Schematic representation of carcinogenesis and progression of bladder cancer. High frequency of LOH in 9q is a common early event and deletion of 2q, 8p and 11q are late events in bladder cancer carcinogenesis. GATA3, FOXA1, and PPARγ are upregulated in the luminal pathway of bladder cancer progression. LOH, loss of heterozygosity; FOXA1, forkhead box A1.

Article Snippet: Membranes were blotted with rabbit polyclonal antibodies against human PPARγ (cat. no. #2430; 1:1,000; Cell Signaling Technology, Inc.), PTEN (cat. no. #9188; 1:1,000; Cell Signaling Technology, Inc.) or α-tubulin (cat. no. PM054-7; 1:5,000; Medical and Biological Laboratories Co., Ltd.) and anti-rabbit IgG horseradish peroxidase-linked antibody (cat. no. #7074; 1:2,000; Cell Signaling Technology, Inc.), according to the manufacturer's instructions.

Techniques: