pparγ agonist troglitazone  (Tocris)

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Promoting APC function of B cells via reprogramming the fatty acid metabolism enhances anticancer immunity in metastatic ovarian cancer

doi: 10.1007/s00262-026-04387-y

Figure Lengend Snippet: The enhanced APC function of B cells by OA in vitro is achieved through H3K27ac-mediated upregulation of PPAR γ expression. A Protein expression of β -actin and H3K27ac in 3 w OvCa-bearing mouse ascitic CD19 + B cells pretreated with BMS and treated with OA was assessed by WB. β -Actin was used as the internal control to calculate the relative expression level of H3K27ac. B The enrichment percentage of H3K27ac at the PPAR γ , CD80, CD86, and CD83 promoter regions was quantified using ChIP-seq analysis. C Correlation analysis of PPAR γ and APC function-related genes (CD80, CD86, CD83, MHC II) in B cells in OvCa patients in the TCGA database (n = 426). D Analysis of protein interaction among FABP4, PPAR γ , CD80, CD86, and CD83. E The enrichment percentage of PPAR γ at the CD80, CD86, and CD83 promoter regions was quantified using ChIP-seq analysis. F Mean fluorescence intensity of CD80, CD86, and CD83 in 3 w OvCa-bearing mouse ascitic CD19 + B cells pretreated with BMS, GW9662/Trog, and treated with OA was detected by flow cytometry. AS, Ascites; Ag, Antigen; OA, oleic acid; BMS, BMS309403. Data are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns, not significant

Article Snippet: In the mechanistic study, mouse ascitic CD19 + B cells (1 × 10 6 /ml) were pretreated with fatty acid binding protein 4 gene (FABP4) inhibitor (BMS309403, MedChemExpress; Cat# HY-101903; 50 μM), PPARγ antagonist (GW9662, MedChemExpress; Cat# HY-16578; 25 μM), and PPAR γ agonist (Troglitazone, Trog, MedChemExpress; Cat# HY-50935; 10 μM) for 2h, respectively.

Techniques: In Vitro, Expressing, Control, ChIP-sequencing, Fluorescence, Flow Cytometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Promoting APC function of B cells via reprogramming the fatty acid metabolism enhances anticancer immunity in metastatic ovarian cancer

doi: 10.1007/s00262-026-04387-y

Figure Lengend Snippet: The mechanism diagram of combining adoptive immunotherapy with APC-function-enhanced B cells and LDC improves anticancer immunity in metastatic OvCa mice. In vitro, OA enhances the expression of CD80/CD86/CD83 of B cells through the H3K27ac-mediated upregulation of PPAR γ expression. Then, the B cells are adoptively transferred into the abdominal cavity of metastatic OvCa mice. In the abdominal cavity, LDC facilitates the APC function of the transferred B cells by causing apoptosis of cancer cells. The transferred B cells promote the activation, proliferation, and differentiation of T cells, which release higher levels of effector molecules (IL-2, GZMB, and IFN- γ , etc.), and facilitate the formation of lymphoid aggregates, finally improving anticancer immunity. LDC, low-dose chemotherapy

Article Snippet: In the mechanistic study, mouse ascitic CD19 + B cells (1 × 10 6 /ml) were pretreated with fatty acid binding protein 4 gene (FABP4) inhibitor (BMS309403, MedChemExpress; Cat# HY-101903; 50 μM), PPARγ antagonist (GW9662, MedChemExpress; Cat# HY-16578; 25 μM), and PPAR γ agonist (Troglitazone, Trog, MedChemExpress; Cat# HY-50935; 10 μM) for 2h, respectively.

Techniques: In Vitro, Expressing, Activation Assay

Journal: iScience

Article Title: Trigonelline activates NRF2 and awakens dormant ovarian follicles to promote pregnancy in aging mice

doi: 10.1016/j.isci.2025.113717

Figure Lengend Snippet: TRG-activated primordial follicles through the NRF2/EGF/AKT signaling pathway (A) Photos of mouse ovaries cultured in medium supplemented with TRG (0.5 μM) and perifosine (100 μM) (TRG+perifosine) at the same time. Scale bars: 200, 100, and 25 μm. (B) Photos of mouse ovary sectioning showing follicles in tissue treated with TRG and perifosine, and histological analysis revealed a decline in the activation of primordial follicles in cultured ovaries from the TRG+perifosine group. Mean ± SE; one-way ANOVA and Tukey’s multiple comparison test; ∗∗∗∗ p < 0.0001, ∗ p = 0.0399, and ∗ p = 0.0138. (C) Levels of P-AKT at the 473-serine site in cultured ovaries from the control, TRG, and TRG+perifosine groups. Mean ± SE; one-way ANOVA, and Tukey’s multiple comparison test; ∗ p = 0.022, ∗∗∗ p = 0.0004, and ∗∗∗∗ p < 0.0001. (D) Levels of P-AKT protein in non-cultured ovaries and ovaries cultured for 6 h from the control and TRG groups. Mean ± SE; one-way ANOVA, and Tukey’s multiple comparison test; ∗p = 0.0153 and ∗p = 0.0406. (E) Levels of FOXO3a protein in ovaries cultured for 7 days from the control and TRG groups. Mean ± SE; two-tailed t test. (F) Levels of P-FOXO3a protein in ovaries cultured for 7 days from the control and TRG groups. Mean ± SE; two-tailed t test. (G) Levels of PTEN protein in ovaries cultured for 7 days from the control and TRG groups. Mean ± SE; two-tailed t test. (H) Levels of MTOR protein in ovaries cultured for 7 days from the control and TRG groups. Mean ± SE; two-tailed t test. (I) Levels and fold change of pS6K/S6K proteins in ovaries cultures for 6 h from the control and TRG groups. (J) Level of EGF protein in cultured ovaries from the control and TRG groups. Mean ± SE; two-tailed t test; ∗ p = 0.0350. (K and L) Level of P-AKT after EGFR inhibition using erlotinib (5 μM and 10 μM) and (L) schematic mode of action of TRG in primordial follicles. TRG induces the nuclear translocation of NRF2 and upregulates NRF-responsive genes, including Egf , leading to increased levels of the EGF protein. EGF then autocrinally activates its receptor, EGFR, leading to phosphorylation and activation of AKT. The activated P-AKT phosphorylates FOXO3a, governing the cytoplasmic localization of FOXO3A and removing its inhibitory effect on cell proliferation, leading to the activation of primordial follicles.

Article Snippet: Ovaries were removed from cervical dislocated F1 (C57Bl/6JRj × CBA/JRj) mice at the indicated ages, randomly separated into control and treatment groups and cultured on inserts (pore size of 0.4 μm, 6.5 mm diameter, Corning, MA, United States) in 24-well plates (Corning, MA, United States) at 37°C in a humidified atmosphere of 5% CO 2 –95% air. α-MEM (Fisher Scientific, Cat# 22571-020 (Gibco)) supplemented with 10% foetal bovine serum (FBS) Fisher Scientific, Cat# 11550356); 5 μg/ml insulin, 5 μg/ml transferrin, and 5 ng/ml sodium selenite (1× ITS; Fisher Scientific, Cat# 12097549); 1× penicillin–streptomycin solution (Thermo Fisher Scientific Cat# 15140122); and 100 mIU/ml rFSH (Merck, Cat# GONAL-F75IE-IU) were used, as described., , Treatment included 0.1-20 μM TRG (Merck, Cat# T5509) (optimal concentration 0.5 μM in saline (TRG)), 1-100 μM perifosine (Selleck Chem Cat# KRX-0401) (optimal concentration is 100 μM) and 5-10 μM erlotinib (Nordic Biosite Cat# 33-A8234) (optimal concentration is 5 μM), as indicated.

Techniques: Cell Culture, Activation Assay, Comparison, Control, Two Tailed Test, Inhibition, Translocation Assay, Phospho-proteomics