Journal: Journal of lipid research

Article Title: Disruption of nucleotide biosynthesis reprograms mitochondrial metabolism to inhibit adipogenesis.

doi: 10.1016/j.jlr.2024.100641

Figure Lengend Snippet: Fig. 4. PPARγ overexpression rescues mitochondrial function induced by the loss of de novo nucleotide biosynthesis. A: OXPHOS protein levels were measured in primary SVF cells that were differentiated and treated with PPARγ inhibitor SR 16832 (2 μM or 10 μM) for 6 days (B) 3T3-L1 cells stably expressing pBABE control vector or PPARγ2 were differentiated and treated with 10 μM 5FU or DMSO (control) for 6 days. Protein expression was analyzed by Western blot as indicated. C: 3T3-L1 cells stably expressing pBABE control vector or PPARγ2 were differentiated and treated with 10 μM MIZ or DMSO (control) for 6 days. Protein expression was analyzed by Western blot as indicated. D: Live cell imaging with MTG and TMRE staining was used to assess changes in mito- chondrial membrane potential relative to mitochondrial mass in the presence or absence of 25 μM MIZ. E: ImageJ was used to measure the ratio of MTG and TMRE. All experiments were repeated two or three times with two or three biological replicates. Statistical significance was determined using one-way ANOVA multiple comparisons test. Error bars indicate mean ± SD, ****P < 0.0001.

Article Snippet: PPARγ2 plasmid (Addgene #8859) or pBabe control was transfected into PLAT-E cells.

Techniques: Over Expression, Stable Transfection, Expressing, Control, Plasmid Preparation, Western Blot, Live Cell Imaging, Staining, Membrane

Journal: Journal of Biological Chemistry

Article Title: Loss of Mitogen-activated Protein Kinase Phosphatase-1 Protects from Hepatic Steatosis by Repression of Cell Death-inducing DNA Fragmentation Factor A (DFFA)-like Effector C (CIDEC)/Fat-specific Protein 27

doi: 10.1074/jbc.m110.210237

Figure Lengend Snippet: FIGURE 5. Increased MAPK activity and PPAR phosphorylation in db/m; mkp-1/ hepatocytes. A, primary hepatocytes isolated from db/m;mkp- 1/ and db/m;mkp-1/ mice and immunoblotted for phospho- and total MAPKs. Phospho-MAPK values were assessed by densitometry and normal- ized to total MAPK values. Data are mean S.E. (error bars; n 8 or 9; *, p 0.05, **, p 0.005). B, quantitative RT-PCR of PPAR mRNA from livers of 8-week-old male db/db;mkp-1/ and db/db;mkp-1/ (left panel) and male mkp-1/ and mkp-1/ mice fed a high fat diet for 16 weeks (right panel). Data are mean S.E. (n 9; *, p 0.05). C, SkHep cells transfected with FLAG-PPAR or FLAG-PPAR S112A with or without MKP-1 and immuno- blotted for Ser(P)112 and total PPAR. Data are representative of three inde- pendent experiments. D, primary hepatocytes from db/m;mkp-1/ or db/m; mkp-1/ mice transfected with pcDNA3 or FLAG-PPAR. FLAG was immunoprecipitated (IP) and immunoblotted for Ser(P)112 and total PPAR (n 3). E, SkHep cells transfected with vector or FLAG-PPAR. MKP-1 was immunoprecipitated and immunoblotted for MKP-1 or PPAR.

Article Snippet: Hepatocytes were transfected with pcDNA3, pcDNA3FLAG-PPAR (Addgene plasmid 8895) (18), pcDNA3FLAG-PPAR S112A, and pcDNA3-MKP-1 with Lipofectamine 2000 (Invitrogen).

Techniques: Activity Assay, Phospho-proteomics, Isolation, Quantitative RT-PCR, Transfection, Immunoprecipitation, Plasmid Preparation

Journal: Open Life Sciences

Article Title: Sini Decoction Intervention on Atherosclerosis via PPARγ-LXRα-ABCA1 Pathway in Rabbits

doi: 10.1515/biol-2018-0053

Figure Lengend Snippet: Expression profiling of PPARγ-LXRα signaling to confirm the mechanism of treating AS by SND. The PPARγ (A-D) and LXRα (E-H) proteins levels in liver, peritoneal macrophages, PMC, and adipose tissue were measured by western blotting. The relative quantities were analyzed by ImageJ software. Data shown as mean ± SEM (n = 6). # P<0.05, compared with Chow group; *P<0.05, compared with HFD group.

Article Snippet: Primary antibodies to ABCA1 and Apo-B were purchased from Abcam (MA, USA); PPARγ antibody and LXRα antibody were purchased from Proteintech (IL, USA); and ApoA-I antibody was purchased from MyBioSource (CA, USA).

Techniques: Expressing, Western Blot, Software

Journal: Advances in Biological Chemistry

Article Title: Piceatannol Enhances Intracellular Energy Metabolism via SIRT1 in C2C12 Cells

doi: 10.4236/abc.2025.153005

Figure Lengend Snippet: Figure 2. PIC enhanced energy metabolism-related genes protein expression. (A) mRNA expression levels of SIRT1, PDK4, PGC1A, and PPARG were determined in C2C12 myo- tubes treated with DMSO, 50 µM, or 100 µM PIC for 24 hours. Results are shown as mean ± SEM (n = 3 - 5). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (B) Protein levels of SIRT1, PDK4, PGC1A, and PPARG were determined in C2C12 myotubes treated with DMSO or 100 µM PIC for 24 hours. Results are shown as mean ± SEM (n = 3 - 5). *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: After blocking with a 4% Block Ace solution (DS Pharma Biomedical, Osaka, Japan) at room temperature for 1 hour, the membranes were incubated overnight at 4 ̊C with a 1:1000 dilution of primary antibodies against SIRT1 (Cell Signaling Technology, Danvers, MA, USA), PGC1A (Proteintech, Rosemont, IL, USA), PDK4 (Abcam, Cambridge, UK), PPARG (Santa Cruz Biotechnology, Santa Cruz, CA, USA), or β-actin (Sigma-Aldrich).

Techniques: Expressing

Journal: Advances in Biological Chemistry

Article Title: Piceatannol Enhances Intracellular Energy Metabolism via SIRT1 in C2C12 Cells

doi: 10.4236/abc.2025.153005

Figure Lengend Snippet: Figure 6. Knockdown of SIRT1 suppressed the PIC-induced upregulation of energy me- tabolism-related gene expression. (A) mRNA levels of SIRT1, PDK4, PGC1A, PPARG in C2C12 myotubes transfected with SIRT1 (+) or control (−) siRNA and then treated with DMSO, 50 µM, or 100 µM PIC for 24 hours. Results are presented as mean ± SEM (n = 4 - 5). *P < 0.05, **P < 0.01, ****P < 0.0001. (B) mRNA levels of CD36, CPT1B, LCAD, and MCAD in C2C12 myotubes transfected with SIRT1 (+) or control (−) siRNA and then treated with DMSO, 50 µM, or 100 µM PIC for 24 hours. Results are presented as mean ± SEM (n = 4 - 5). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: After blocking with a 4% Block Ace solution (DS Pharma Biomedical, Osaka, Japan) at room temperature for 1 hour, the membranes were incubated overnight at 4 ̊C with a 1:1000 dilution of primary antibodies against SIRT1 (Cell Signaling Technology, Danvers, MA, USA), PGC1A (Proteintech, Rosemont, IL, USA), PDK4 (Abcam, Cambridge, UK), PPARG (Santa Cruz Biotechnology, Santa Cruz, CA, USA), or β-actin (Sigma-Aldrich).

Techniques: Knockdown, Gene Expression, Transfection, Control

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: Upregulation of NCoA6 expression is correlated with PDAC progression. (A) NCoA6 expression is significantly increased in PDAC samples compared with normal tissues from TCGA and GTEx. (B) Representative images of IHC staining for NCoA6 in PDAC tumor tissues (T) and adjacent normal tissues (ANT). (C) IHC score of NCoA6 expression in PDAC and ANT ( n = 55, p < 0.001). (D) Representative images of IHC staining for NCoA6 in tissue microarrays. (E) Kaplan–Meier analysis of NCoA6 expression and overall survival ( p < 0.01) and disease‐free survival ( p < 0.05) of patients with PDAC from FUSCC. (F) The high NCoA6 expression group is associated with worse overall and disease‐free survival according to the Kaplan–Meier plotter database (hazard ratio [HR] = 1.44 and 3.15, respectively). (G) Time‐dependent ROC curve shows that NCoA6 expression in PDAC tissues from FUSCC is related to the 2‐year overall and disease‐free survival of patients (AUC = 0.822 and 0.707, respectively). (H) The ROC analysis from UCSC shows the moderate prognostic value of NCoA6 expression level in PDAC tissues (AUC = 0.811, p < 0.001).

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Expressing, Immunohistochemistry

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: Relationship between clinicopathological features and NCoA6 expression in patients with pancreatic cancer.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Expressing

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: Univariate and multivariate Cox regression of overall survival and disease‐free survival of patients with PDAC.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Expressing

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: GO/KEGG analysis and GSEA of our RNA‐sequencing data. (A) Western blotting analysis of NCoA6 in PDAC lines. The H6C7 cell line was included as a negative control for the detection of endogenous NCoA6 expression, and β‐Actin was used as a control. (B) RT‐qPCR and (C) western blotting analysis of NCoA6 expression in NCoA6 PANC‐1 and SW1900 cells. The volcano plot (D) and heatmap (E) of gene alterations for NCoA6‐NC/sh PANC‐1 cells. (F) GSEA shows that SENESE_HDAC1_TARGETS_UP, the gene set from Human MSigDB Collections, is significantly enriched in the RNA‐sequencing dataset of NCoA6‐NC/sh PANC‐1 cells. (G) The main enriched GO/KEGG results of NCoA6‐NC/sh PANC‐1 cells.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: RNA Sequencing, Western Blot, Negative Control, Expressing, Control, Quantitative RT-PCR

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: NCoA6 knockdown decreases the proliferation, cell cycle, and apoptosis of pancreatic cancer cells. CCK‐8 assays (A, B) and colony formation assays (C, D) were used to test the proliferation of PDAC cells transfected with NCoA6 shRNAs. The representative images and statistical results of the cell cycle (E, F) and apoptosis (G, H) using flow cytometry. (I, J) Western blotting analysis of p21 and p27 expression in NCoA6‐silenced PANC‐1 and SW1990 cells. (K, L) Western blotting analysis of CDK4, CDK2, Cyclin D1, Cyclin E1, and Cyclin A2 expression in NCoA6‐silenced PANC‐1 and SW1990 cells.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Knockdown, CCK-8 Assay, Transfection, Flow Cytometry, Western Blot, Expressing

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: NCoA6 knockdown decreases the migration and invasion capacity of pancreatic cancer cells. Scratch assays (A, B) and transwell migration and invasion assays (C, D) were performed to assess the effect of NCoA6 silencing on pancreatic cancer cell lines. (E, F) Western blotting analysis of EMT protein expression in NCoA6‐silenced PANC‐1 and SW1990 cells.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Knockdown, Migration, Western Blot, Expressing

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: Potential endogenous interactions of NCoA6, HDAC1, FBW7, and CDX2. (A, B) Co‐immunoprecipitation validated NCoA6 interaction with HDAC1 in PDAC cells. NCoA6 expression was negatively correlated with FBW7 and CDX2 expression. (C) Western blotting validated NCoA6 downregulation in FBW7 overexpressing PANC‐1 and SW1990 cells. (D, E) Correlation analysis of NCoA6 expression and FBW7 expression in PDAC tissues, as determined by the IHC score (*** p < 0.001). (F, G) Western blotting and RT‐qPCR validated CDX2 upregulation in NCoA6 knockdown PDAC cells. (H, I) Western blotting and RT‐qPCR validated CDX2 upregulation in HDAC1 knockdown PDAC cells. (J) AcH3, AcH4, and CDX2 were upregulated in HDAC inhibitor TSA‐treated PDAC cells (western blotting).

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Immunoprecipitation, Expressing, Western Blot, Quantitative RT-PCR, Knockdown

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: Schematic representation of the working model designed using FigDraw. NCoA6 plays an important role in promoting PDAC cell proliferation, migration, invasion and progression, and is potentially related to the expression of HDAC1, FBW7, and CDX2.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Migration, Expressing