Journal: Frontiers in Immunology

Article Title: miR-223 alleviates DSS-induced colitis by prompting macrophage M2 polarization through PPAR-γ/FOXO1 signaling

doi: 10.3389/fimmu.2025.1598781

Figure Lengend Snippet: Relieving colitis by miR-223 through the promotion of macrophage M2 polarization via the modulation of PPAR-γ/FOXO1 signaling. (A) Representative images of western blots indicating the protein expression levels of PPAR-γ and FOXO1. The data represent the findings from three independent experiments. (B, C) Changes in the expression of PPAR-γ and FOXO1. (D, E) Changes in the mRNA levels of PPAR-γ and FOXO1. All the data are expressed as the means ± SDs (n=6 each group). *p<0.05, **p<0.01, ***p<0.001. (F, G) Correlations of colonic miR-223 expression with PPAR-γ and FOXO1 expression in the colon. Pearson analyses were used to correlate miR-223 with colonic PPAR-γ and FOXO1 expression. PPAR-γ, peroxisome proliferator-activated receptor gamma; FOXO1, Forkhead box transcription factor O1.

Article Snippet: After being blocked with bovine serum albumin blocking buffer (3% in PBS), the slides were incubated with the following primary antibodies: anti-PPAR-γ antibody (1:3000) (Cat.# GB11163, Servicebio, Wuhai, China), anti-FOXO1 antibody (1:10000) (Cat.# GB11286-1, Servicebio), anti-CD206 (1:5000) (Cat.# GB113497 , Servicebio), anti-CD86 (1:5000) (Cat.# GB115630 , Servicebio) or anti-F4/80 (1:2000) (Cat.# GB113373 , Servicebio) overnight at 4°C.

Techniques: Western Blot, Expressing

Journal: Frontiers in Immunology

Article Title: miR-223 alleviates DSS-induced colitis by prompting macrophage M2 polarization through PPAR-γ/FOXO1 signaling

doi: 10.3389/fimmu.2025.1598781

Figure Lengend Snippet: Immunofluorescence analysis of PPAR-γ and M2 macrophage marker CD206 in colonic tissue. Representative micrographs depict protein expression levels of PPAR-γ (red) and its co-localization (yellow) with CD206 (green) in colon sections. Nuclei were counterstained with DAPI (blue). Scale bar: 100 µm.

Article Snippet: After being blocked with bovine serum albumin blocking buffer (3% in PBS), the slides were incubated with the following primary antibodies: anti-PPAR-γ antibody (1:3000) (Cat.# GB11163, Servicebio, Wuhai, China), anti-FOXO1 antibody (1:10000) (Cat.# GB11286-1, Servicebio), anti-CD206 (1:5000) (Cat.# GB113497 , Servicebio), anti-CD86 (1:5000) (Cat.# GB115630 , Servicebio) or anti-F4/80 (1:2000) (Cat.# GB113373 , Servicebio) overnight at 4°C.

Techniques: Immunofluorescence, Marker, Expressing

Journal: Frontiers in Immunology

Article Title: miR-223 alleviates DSS-induced colitis by prompting macrophage M2 polarization through PPAR-γ/FOXO1 signaling

doi: 10.3389/fimmu.2025.1598781

Figure Lengend Snippet: Schematic representation of the potential mechanisms underlying the targeted therapy of miR-223 supplement for DSS-induced colitis. miR-223 ameliorates DSS-induced colitis through promoting macrophage M2 polarization via modulation of PPAR-γ and FOXO1 signaling. DSS, dextran sodium sulfate.

Article Snippet: After being blocked with bovine serum albumin blocking buffer (3% in PBS), the slides were incubated with the following primary antibodies: anti-PPAR-γ antibody (1:3000) (Cat.# GB11163, Servicebio, Wuhai, China), anti-FOXO1 antibody (1:10000) (Cat.# GB11286-1, Servicebio), anti-CD206 (1:5000) (Cat.# GB113497 , Servicebio), anti-CD86 (1:5000) (Cat.# GB115630 , Servicebio) or anti-F4/80 (1:2000) (Cat.# GB113373 , Servicebio) overnight at 4°C.

Techniques:

Journal: Journal of Lipid Research

Article Title: Adipocyte expression of the glucose-dependent insulinotropic polypeptide receptor involves gene regulation by PPARγ and histone acetylation

doi: 10.1194/jlr.m012203

Figure Lengend Snippet: Fig. 7. RNAi-mediated suppression of PPAR reduces adipocyte GIPR expression. A: Effects of PPAR siRNAs on PPAR expression are shown. Differentiated 3T3-L1 adipocytes were transfected with a pool of three siRNAs for PPAR or control scrambled siRNAs (100 nM) and incubated for 72 h. Nuclear extracts were isolated, and Western blot analyses were performed as described in “Experi- mental Procedures,” using antibodies against PPAR 2, PPAR , and histone H3. B: Effects of PPAR siRNAs on GIPR expression are shown. 3T3-L1 adipocytes were treated as described in the legend to panel A, and real-time RT-PCR was performed with extracts to quantify GIPR mRNA levels, shown as the fold difference versus control normalized to 18S rRNA expression levels. All data repre- sent three independent experiments, each carried out in duplicate, and Western blots are representative of n = 3 replicates. Signifi - cance was tested using a Student t test, where ** represents P <0.05 versus control siRNA.

Article Snippet: Differentiated 3T3-L1 adipocytes were transfected with a pool of three small interfering RNAs (siRNAs) for PPAR (sc-29456; Santa Cruz Biotechnology), using Lipofectamine 2000 transfection reagent, and incubated for 72 h. The reduction in PPAR expression level was determined by Western blot hybridization using antibody against PPAR 2 (sc-22020; Santa Cruz Biotechnology). at N O R T H D A K O T A S T A T E U N IV , on M ay 30, 2015 w w w .jlr.org D ow nloaded from increase in GIPR mRNA levels was detected (see Fig. 2D, treatments 4 and 5).

Techniques: Expressing, Transfection, Control, Incubation, Isolation, Western Blot, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: Transcriptional Regulation of Flotillins by the Extracellularly Regulated Kinases and Retinoid X Receptor Complexes

doi: 10.1371/journal.pone.0045514

Figure Lengend Snippet: Oligonucleotides and plasmids.

Article Snippet: pSV Sport PPARγ , PPARγ expression plasmid , Addgene 8886, .

Techniques: Luciferase, Plasmid Preparation, Expressing, Control, Dominant Negative Mutation

Journal: PLoS ONE

Article Title: Transcriptional Regulation of Flotillins by the Extracellularly Regulated Kinases and Retinoid X Receptor Complexes

doi: 10.1371/journal.pone.0045514

Figure Lengend Snippet: Flotillin promoter constructs F1-1330 (A, C) or F2-2130 (B, D) were cotransfected into Hela cells together with expression plasmids for RAR, RXR, PPARγ or with empty PSV control plasmid. One day post-transfection, the cells were stimulated with trans-RA (1 µM) for 24 h in serum-free medium. Relative luciferase activity of the unstimulated control sample was set as 1. F1-1330 (E) and F2-2130 (F) transfected Hela cells were stimulated with troglitazone for 24 h in serum-free medium. Values are mean ± standard deviation of at least 3 experiments measured in duplicates. ***p<0.001; **p<0.01; *p<0.05 vs. respective control.

Article Snippet: pSV Sport PPARγ , PPARγ expression plasmid , Addgene 8886, .

Techniques: Construct, Expressing, Control, Plasmid Preparation, Transfection, Luciferase, Activity Assay, Standard Deviation

Journal: PLoS ONE

Article Title: Transcriptional Regulation of Flotillins by the Extracellularly Regulated Kinases and Retinoid X Receptor Complexes

doi: 10.1371/journal.pone.0045514

Figure Lengend Snippet: Hela cells were transiently transfected with expression constructs for RAR, RXR, or a combination of both. Empty PSV vector served as a control. One day post-transfection, the cells were stimulated with trans-RA (1 µM) in serum-free medium for 24 h. Cell lysates were analyzed for flotillin-1 (A), flotillin-2 (B), RAR and RXR (C) by Western blotting. D and E show a densitometric quantification of flotillin expression. F: Cells were transfected with RAR or PPARγ expression construct or empty PSV. RNA was isolated, transcribed into cDNA and flotillin mRNA was measured by qPCR. Values are mean ± standard deviation of at least 3 experiments. ###, p<0.001; #, p<0.05; vs control *, p<0.05 vs. unstimulated sample.

Article Snippet: pSV Sport PPARγ , PPARγ expression plasmid , Addgene 8886, .

Techniques: Transfection, Expressing, Construct, Plasmid Preparation, Control, Western Blot, Isolation, Standard Deviation

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

Journal: Pharmaceuticals

Article Title: Saroglitazar Mitigated Cyclophosphamide-Induced Testicular Injury: Crosstalk Between Oxidative Stress, Inflammation and Apoptosis

doi: 10.3390/ph19020266

Figure Lengend Snippet: Impact of SAR2 and SAR4 on CYC-induced changes in peroxisome proliferator-activated receptor gamma (PPAR-γ), nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1). ( A ) Testicular PPAR-γ level, ( B ) Testicular Nrf2 level, ( C ) Testicular HO-1 level. CYC: cyclophosphamide, SAR: saroglitazar. Data are expressed as mean ± S.E. One-way ANOVA followed by post hoc Tukey’s multiple comparison test was used for comparison of mean values. * p < 0.05, vs. control group, # p < 0.05 vs. CYC group, $ p < 0.05 vs. SAR 2 group.

Article Snippet: Testicular homogenate was used for determination of TNF-α (Elabscience, Wuhan, China, Cat. no. E0764Ra), IL-6 (Cloud-Clone Corp, Wuhan, China, Cat. no. SEA079Ra), Bcl2 (Elabscience, China, Cat. no. ER0762), Bax (Elabscience, China, Cat. no. ERO512), PPAR-γ (Elabscience, China, Cat. no. E-EL-R0724), Nrf2 (RayBiotech, Peachtree Corners, GA, USA, Cat. no. RD-NFE2L2-Ra) and HO-1 (CUSABIO, Wuhan, China, CSB-E08267r) via ELISA as instructed by the manufacturer.

Techniques: Comparison, Control