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: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Sex hormone binding globulin (SHBG) modulates mitochondrial dynamics in PPARγ-depleted equine adipose derived stromal cells

doi: 10.1007/s00109-024-02459-z

Figure Lengend Snippet: The antibodies used for Western Blot

Article Snippet: PPARG , Aviva Systems Biology, ARP32880 , 1:500.

Techniques: Western Blot

Journal: Journal of medicinal chemistry

Article Title: A novel N-substituted valine derivative with unique PPARγ binding properties and biological activities

doi: 10.1021/acs.jmedchem.0c01555

Figure Lengend Snippet: Effect of 7j on PPARγ phosphorylation. (A) Percentage of in vitro PPARγ Ser273 phosphorylation by CDK5 in the presence of 0.1 μM of 7j or Rosi. (B) Phosphorylation of PPARγ Ser273 in 3T3-L1 adipocyte incubated for 60 minutes with 5 μM of Rosi, 7j or Roscovitine (Rsv) before TNFα stimulation (50 ng/ml; 90 minutes). Two independent experiments are shown. Phosphospecific antibodies were from Rockland or New England Peptide (NEP). Arrow heads indicate phosphorylated PPARγ and PPARγ2. (C) RT-PCR analysis of the expression levels of a selection of genes known to be regulated by CDK5-dependent phosphorylation of PPARγ. Values are means ± SD (n = 5) expressed relative to the mean of control. **p < 0.01, ***p < 0.001 vs. control (one-way ANOVA followed by Dunnett’s post hoc test).

Article Snippet: PPARγ phospho Ser273 antibodies were from Rockland (Limerick, PA, USA) or custom produced by New England Peptide (Gardner, MA, USA).

Techniques: Phospho-proteomics, In Vitro, Incubation, Reverse Transcription Polymerase Chain Reaction, Expressing, Selection, Control