Journal: Archives of Toxicology

Article Title: Bisphenol Z at environmentally relevant dose dysregulates mitochondrial metabolism and proliferative capacity in human ovarian granulosa cells by inducing changes in metabolic substrate utilization

doi: 10.1007/s00204-025-04278-8

Figure Lengend Snippet: BPZ disrupts fatty acids utilisation in human ovarian GCs. Schematic representation of fatty acids utilization and inhibitory action of Etomoxir ( A ). Oxygen consumption rate (OCR) changes post BPZ treatment in fatty acids dependency ( B ) and capacity ( C ) tests. BPZ-induced changes in fatty acids dependency, capacity and flexibility in primary GCs ( D , E , F ). Effects of BPZ on mRNA expression of fatty acids transporters, SLC27A1 ( G ) and CD36 ( H ) in HGrC1 cells. PPARA ( I ), PPARD ( J ) and PPARG ( K ) mRNA expression after BPZ treatment in primary GCs. Fatty acids uptake in primary GCs after 6 h BPZ treatment ( L ). Changes in viability post BPZ, fatty acids dependent pathway inhibitor, Etomoxir and co-treatment with both in HGrC1 cells ( M ). C, control (0.1% DMSO). Data are mean ± SD of three independent experiments. ** p ≤ 0.01; control vs. experimental group ( D – I ). Statistically significant are the mean values not sharing letters, p ≤ 0.05; pairwise comparison between each experimental group and the control ( J )

Article Snippet: To assess the mRNA expression of glucose, fatty acids and glutamine transporters real-time PCR was performed using following genes: glucose transporter 1 (GLUT1, SLC2A1 ; Hs00892681_m1), glucose transporter 4 (GLUT4, SLC2A4 ; Hs00168966_m1), CD36 molecule ( CD36 , hs00354519_m1), solute carrier family 27 member 1 ( SLC27A1 , hs01587911_m1), solute carrier family 1 member 5 ( SLC5A1 , Hs01056542_m1) and peroxisome proliferator‐activated receptors: α ( PPARA , Hs00947536_m1), δ ( PPARD , Hs04187066_g1) and γ ( PPARG , Hs00234592_m1).

Techniques: Expressing, Control, Comparison

Journal: Journal of translational medicine

Article Title: An integrated approach of network pharmacology, molecular docking, and experimental verification uncovers kaempferol as the effective modulator of HSD17B1 for treatment of endometrial cancer.

doi: 10.1186/s12967-023-04048-z

Figure Lengend Snippet: Fig. 5 Kaempferol modulated estrogen metabolism pathways and differentially regulates PPARG expression in EC cells of different ER subtypes. A– B HSD17B1 and HSD17B1-associated genes, such as ESRRA, PPARG, and ESR1, are involved in several estrogen metabolism pathways, such as steroid binding, 17- beta-hydroxysteroid dehydrogenase (NADP+) activity, steroid hormone biosynthesis, and regulation of hormone levels. C Kaempferol suppressed the expression of PPARG in ER-positive AN3 CA and promoted the expression of PPARG in ER-negative HEC-1-A. D–I Kaempferol suppressed the expression of PPARGC1A and ESRRA in both AN3 CA (D–F) and HEC-1-A cells (G–I), without modulating ESR1. Western blotting (D–E and G–H) and the IHC scores (F and I) confirmed the differential expression of PPARGC1A and ESRRA. Results are presented as means and SDs. Compared with the negative control, *, #P < 0.05, **, ##P < 0.01, ***, ###P < 0.001

Article Snippet: The whole cell lysates and tumor homogenates (50 μg) were resolved on an 8 ~ 12% SDS–polyacrylamide gel, transferred to a polyvinylidene difluoride membrane (NEN Life Sciences, Boston, MA), probed sequentially with antibodies against ESR1 (ab108398, 67 kDa), ESRRA (ab137489, 55 kDa), PPARGC1A (ab188102, 91 kDa) (Abcam, Cambridge, MA, U. S.), CASP3/p17/p19 (19677–1, 35 kDa), CASP9/p35/p10 (66169–1, 46 kDa), PPARG (16643–1, 55 kDa), HSD17B1 (25334–1, 35 kDa) (Proteintech, Wuhan, China) at 4 °C overnight, rinsed, and incubated with the goat anti-rabbit secondary antibody (Abcam, Cambridge, MA).

Techniques: Expressing, Binding Assay, Activity Assay, Western Blot, Quantitative Proteomics, Negative Control

Journal: Cells

Article Title: RXR Agonists Enhance Lenalidomide Anti-Myeloma Activity and T Cell Functions while Retaining Glucose-Lowering Effect.

doi: 10.3390/cells12151993

Figure Lengend Snippet: Figure 3. LG100754 attenuates the binding effect of PPARα and PPAR γ on the CRBN promoter area. (A) U266 and MM1.R were transfected with CRBN/PGL3 firefly luciferase reported vector construct, then co-treated with PPARs agonist with LG100754 for 48 h, and luciferase bio-luminate activity was measured. (B) Bar graphs show qRT-PCR data using immunoprecipitated DNA obtained from ChIP with anti-CRBN or anti-IgG (negative control) antibodies; error bars represent SD. Results are presented as mean ± SD from at least three separate experiments. NS: not statistically significant; *: p < 0.05; **: p < 0.01.

Article Snippet: The PPARβ/δ (NBP2-22468) and PPARγ (NBP2-22106) antibodies were obtained from Novus Biologicals, LLC. (Littleton, CO, USA).

Techniques: Binding Assay, Transfection, Luciferase, Plasmid Preparation, Construct, Activity Assay, Quantitative RT-PCR, Immunoprecipitation, Negative Control

Journal: Free radical biology & medicine

Article Title: Redox-dependent PPARγ/Tnpo1 complex formation enhances PPARγ nuclear localization and signaling.

doi: 10.1016/j.freeradbiomed.2020.06.005

Figure Lengend Snippet: Fig. 1. PPARγ2 and Tnpo1 bind via disulfide bonds. (A–C) Six-week-old male C57BL/6 mice were intravenously injected via the tail vein with 4 × 109 plaque-forming units of mock adenovirus or PPARγ2 adenovirus (adPPARγ2). Livers were dissected 2 weeks later for analysis (n = 3/group). (A) Hepatic TG levels. **P < 0.01 vs. mice infected with mock adenovirus. (B) Immunoprecipitation of PPARγ2/Tnpo1, PPARγ2/importin-α, and PPARγ2/importin-β complexes from liver samples. (C) Immunoprecipitation of PPARγ2/Tnpo1 complex from liver samples with or without 10 mM DTT treatment. (D–F) Immunoprecipitation of PPARγ2:ΔCys (Ser→Cys)/Tnpo1 (D), PPARγ2:WT (Cys→Ser)/ Tnpo1 (E), and PPARγ2/Tnpo1:WT (Cys→Ser) complexes (F). (G) PPARγ2/Tnpo1 complex (yellow), PPARγ2 (green), and Tnpo1 (red) in mouse primary cultured hepatocytes transfected with PPARγ2 or PPARγ2:ΔCys expression vector for 48 h, then treated with 250 μM H2O2 or left untreated for 6 h. Cell lysates were immunoprecipitated with anti-PPARγ antibody, treated with 10 mM DTT or left untreated, and then subjected to non-reducing sodium dodecyl sulphate–polya- crylamide gel electrophoresis followed by western blotting. All data are expressed as mean ± SEM. P value was calculated using unpaired Student's t-test in A. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Mutant mouse Tnpo1 and PPARγ2 expression vectors were prepared using the KOD-Plus-Mutagenesis kit (Toyobo, Osaka, Japan) from MycDDK-tagged mouse Tnpo1 and PPARγ2 expression vectors (Origene).

Techniques: Injection, Infection, Immunoprecipitation, Cell Culture, Transfection, Expressing, Plasmid Preparation, Nucleic Acid Electrophoresis, Western Blot

Journal: Free radical biology & medicine

Article Title: Redox-dependent PPARγ/Tnpo1 complex formation enhances PPARγ nuclear localization and signaling.

doi: 10.1016/j.freeradbiomed.2020.06.005

Figure Lengend Snippet: Fig. 3. Cytosolic H2O2/Tnpo1-dependent nuclear translocation of PPARγ2 enhances PPARγ2 target gene expression and TG accumulation in hepatocytes. (A–C) WT and Sod1-deficient hepatocytes were infected with mock adenovirus or adPPARγ2 at a MOI of 50 for 48 h, followed by culturing with or without 5 μM DMNQ for 6 h. (D–F) WT hepatocytes were infected with mock adenovirus or adPPARγ2 at an MOI of 50, transfected with Tnpo1 or control siRNA for 24 h, and cultured with or without 5 μM DMNQ for 6 h. (A, D) (Left) Oil Red O staining. (Right) Quantification of Oil Red O staining. (B, C, E, F) Quantification of Pparg2, Fsp27, Fabp1, and Fabp4 mRNA levels (B, E) and the amount of DNA-bound PPARγ (C, F) in WT and Sod1-deficient hepatocytes (n = 5/group). *P < 0.05, **P < 0.01 vs. WT hepatocytes infected with mock adenovirus without DMNQ treatment (A–C) and vs. hepatocytes infected with mock adenovirus, transfected with control siRNA, and without DMNQ treatment (D–F). All data are expressed as mean ± SEM. P value was calculated by one-way ANOVA in A, B, C, D, E, and F. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Mutant mouse Tnpo1 and PPARγ2 expression vectors were prepared using the KOD-Plus-Mutagenesis kit (Toyobo, Osaka, Japan) from MycDDK-tagged mouse Tnpo1 and PPARγ2 expression vectors (Origene).

Techniques: Translocation Assay, Targeted Gene Expression, Infection, Transfection, Control, Cell Culture, Staining

Journal: Free radical biology & medicine

Article Title: Redox-dependent PPARγ/Tnpo1 complex formation enhances PPARγ nuclear localization and signaling.

doi: 10.1016/j.freeradbiomed.2020.06.005

Figure Lengend Snippet: Fig. 4. Cytosolic H2O2/Tnpo1-dependent nuclear translocation of PPARγ2 promotes hepatic TG accumulation in mice infected with adPPARγ2. (A–F) Six-week-old male C57BL/6 WT and Sod1-deficient mice were intravenously injected via the tail vein with 4 × 109 plaque-forming units of mock adenovirus and PPARγ2 and SOD1 adenoviruses (adPPARγ2 and adSOD1, respectively) in various combinations. Livers were dissected from mice 2 weeks later for analysis. (G–J) Six-week-old male C57BL/6 WT and Sod1-deficient mice were intravenously injected via the tail vein with 4 × 109 plaque-forming units of mock adenovirus or adPPARγ2, followed by injection of control or Tnpo1 siRNA (1 mg/kg body weight) after 2 days. Livers were dissected from mice 12 days later for analysis. (A, G) Representative H&E-stained liver sections. (F) Immunoprecipitation of PPARγ2/Tnpo1 complex from liver tissue. (B–E, H–J) Quantification of hepatic TG (B, H), cytosolic H2O2 (C), and Pparg2, Fsp27, Fabp1, and Fabp4 mRNA (D, I) levels and the amount of DNA-bound PPARγ (E, J) (n = 5/group). **P < 0.01 vs. WT mice infected with mock adenovirus (B–E); and *P < 0.05, **P < 0.01 vs. WT mice infected with mock adenovirus and treated with control siRNA (H–J). All data are expressed as mean ± SEM. P value was calculated by one-way ANOVA in B, C, D, E, H, I, and J.

Article Snippet: Mutant mouse Tnpo1 and PPARγ2 expression vectors were prepared using the KOD-Plus-Mutagenesis kit (Toyobo, Osaka, Japan) from MycDDK-tagged mouse Tnpo1 and PPARγ2 expression vectors (Origene).

Techniques: Translocation Assay, Infection, Injection, Control, Staining, Immunoprecipitation

Journal: Free radical biology & medicine

Article Title: Redox-dependent PPARγ/Tnpo1 complex formation enhances PPARγ nuclear localization and signaling.

doi: 10.1016/j.freeradbiomed.2020.06.005

Figure Lengend Snippet: Fig. 5. Cytosolic H2O2/Tnpo1-dependent nuclear translocation of PPARγ2 promotes hepatic TG accumulation in mice fed an HFD. (A–F) Six-week-old male C57BL/6 mice were fed a control diet or HFD for 4 weeks, then intravenously injected via the tail vein with control, Tnpo1, Sod1, or Pparg2 siRNA (1 mg/kg body weight) in various combinations. Livers were dissected from the mice 10 days later for analysis (n = 5/group). (A) Representative H&E-stained liver sections. (B–D) Quantification of TG (B), cytosolic H2O2 (C), and Pparg2, Fsp27, Fabp1, and Fabp4 mRNA levels (D) in the liver (n = 5/group). **P < 0.01 vs. mice fed a control diet and treated with control siRNA. (E) Immunoprecipitation of PPARγ2/Tnpo1 complex in the liver. (F) Quantification of the amount of DNA- bound PPARγ in the liver (n = 5/group). **P < 0.01 vs. mice fed a control diet and treated with control siRNA. #P < 0.05, ##P < 0.01 vs. mice fed an HFD diet and treated with control siRNA. All data are expressed as mean ± SEM. P value was calculated by one-way ANOVA in B, C, D, E, and F.

Article Snippet: Mutant mouse Tnpo1 and PPARγ2 expression vectors were prepared using the KOD-Plus-Mutagenesis kit (Toyobo, Osaka, Japan) from MycDDK-tagged mouse Tnpo1 and PPARγ2 expression vectors (Origene).

Techniques: Translocation Assay, Control, Injection, Staining, Immunoprecipitation