anti free fatty acid receptor ffar  (Proteintech)

Journal: Life Metabolism

Article Title: Propionic acid secreted by Akkermansia muciniphila alleviates hepatic fibrosis by antioxidant regulation across the gut–liver axis

doi: 10.1093/lifemeta/loaf036

Figure Lengend Snippet: Propionic acid inhibits the activation of HSCs and oxidative stress. (a) Immunofluorescence analysis (Scale bar = 50 μm). (b) ROS visualized by microscopy using the 2’,7’-dichlorodihydrofluorescein diacetate (DCFH-DA) probe at a final concentration of 10 μmol/L (Scale bar = 50 μm). (c) Fibrosis-related markers such as TIMP-1, α-SMA, and Col1α1 and TGF-β1 protein expression assayed by western blotting. (d) Oxidative stress-related Keap1/Nrf2 pathway protein levels assayed by western blotting. (e–h) MDA level (e), CAT activity (f), SOD activity (g), and GSH-Px activity (h). (i) In vitro experiments conducted to assess the effects of TGF-β, GPR41 inhibitor (SHB), GPR43 inhibitor (GLPG0974), and propionic acid pretreatment on oxidative stress-related Keap1/Nrf2 signaling pathway in LX2 cells through immunoblot analysis. (j) Western blot analysis of oxidative stress-related Keap1/Nrf2 signaling pathway performed in LX2 cells transfected with sh GPR41 , followed by stimulation with TGF-β and pretreatment with propionic acid. Data are presented as the mean ± SEM. Statistical significance was determined using ANOVA for multiple-group comparisons. ** P < 0.01, *** P < 0.05, compared with the control group. # P < 0.05, ## P < 0.01, compared with the model group.

Article Snippet: The study utilized various reagents and kits, including sodium propionate (MedChemExpress, HY-B1773A), 4’,6-diamidino-2-phenylindole (DAPI; Thermo, D3571), MDA, CAT, SOD, and GSH-Px assay kits (Thermo, EEA015; Jianglai Biotechnology, JL18163, JL12237, JL-T0879, respectively), propionic acid ELISA kit (Zhuocai Biotechnology, ZC-56375-J), and antibodies such as Col1α1 (Abcam, RRID: AB_138492), α-SMA (Abcam, RRID: AB_5694), TGF-β1 (Proteintech, 21898-1-AP), SMAD2, p-SMAD2, SMAD3, p-SMAD3 (Cell Signaling Technology, 5339, 18338, 9523, 9520, respectively), CAT (Atlas Antibodies, HPA051282), SOD (Fitzgerald, 70R-13887), Nrf-2 (Cell Signaling Technology, 80593-1-RR), Keap1 (Proteintech, 80744-1-RR), GPR41 (Proteintech, 66811-1-Ig), GPR43 (Proteintech, 19952-1-AP), and Trizol reagent (Beyotime, R0016).

Techniques: Activation Assay, Immunofluorescence, Microscopy, Concentration Assay, Expressing, Western Blot, Activity Assay, In Vitro, Transfection, Control

Journal: Nature Communications

Article Title: Genetic diversity of Collaborative Cross mice implicates FFAR3 as a target for ILC2 anti-inflammatory reprogramming

doi: 10.1038/s41467-025-67813-2

Figure Lengend Snippet: A ILC2s from lungs of C57BL/6J and CC030 after 4 days of culture with IL-33, TSLP, and IL-2 (n = 3 per strain) ** p = 0.0016. B CFSE dilution between ILC2s after 48 h of culture with IL-33, TSLP, and IL-2. C Proliferation index and D division index after 48 h culture as in ( B ) (n = 4 per strain) **** p < 0.0001. E IL-13 in supernatant of ILC2s cultured per (A) (n = 3 per strain) ** p = 0.0018. F IL-13 expression by intracellular cytokine staining. MFI = mean fluorescence intensity (n = 3 per strain) * p = 0.011. G Representative flow plots of active caspase+ ILC2s, pre-gated on live cells (numbers above gates = %). H Quantification of active caspase+ ILC2s as in (G) (n = 4 per strain) *** p = 0.0004. I Representative flow plot of early/late apoptosis by Annexin V and propidium iodide (PI) staining on ILC2s 6 days post culture. J Early and late apoptotic K ILC2s (n = 5 per strain) *** p = 0.0004, ** p = 0.0019. L Apoptotic pathways enriched in naïve C57BL/6J vs CC030 ILC2s. M ILC2s as % of live lung cells and N as % of innate lymphocytes (Lineage⁻, CD90⁺) (n = 3 per strain) ** p = 0.0027, **** p < 0.0001. O IL-5 and P IL-13 in lung homogenate after 4 days of Alt Ex challenge (n = 4 C57BL/6J, 5 CC030) *** p = 0.0006, **** p < 0.0001. Q Total eosinophils in bronchoalveolar lavage (BAL) fluid after 4 days of Alt Ex challenge (n = 5 per strain, also in Supplementary Fig. C) ** p = 0.0027. R Diagram implicating candidate genes in Ch.7 ILC2 locus. S Ffar3 expression by normalized counts from bulk RNA-seq (DESeq2 DGE analysis, Wald test (two-tailed) with Benjamini–Hochberg correction; **** p = 1.76 × 10 −12 ). T qPCR for Ffar3 from RNA of flow-sorted naïve CC030 ILC2s from various tissues (n = 5 mice; one sample each from bone marrow, adipose, and colon had insufficient RNA). A , C – F , H , J , K , M – Q , S , T : bar height = mean, error bars = SD. A , C – F , M – P , S – T : each representative of two independent experiments, H , J , K , Q , S , T : each representative of one independent experiment. Significance for ( A ), ( C – F ), ( H ), ( J , K ), ( L – Q ), assessed by two-tailed unpaired Student’s t -test; for ( T ) by one-way ANOVA with Tukey multiple comparisons (two-tailed).

Article Snippet: Quantitative real-time PCR was carried out with TaqMan Gene Expression assays: Gapdh (Mm99999915) and Ffar3 (Mm02621638_s1) on a QuantStudio 5 Real-Time PCR System (Applied Biosystems) instrument for 45 cycles.

Techniques: Cell Culture, Expressing, Staining, Fluorescence, RNA Sequencing, Two Tailed Test

Journal: Nature Communications

Article Title: Genetic diversity of Collaborative Cross mice implicates FFAR3 as a target for ILC2 anti-inflammatory reprogramming

doi: 10.1038/s41467-025-67813-2

Figure Lengend Snippet: A Schematic for vehicle- vs. AR420626 (10 µM)–treated ILC2s by bulk RNA-sequencing. B Volcano plot of differentially expressed genes from ( A ). Egfr encircled. Labels = 10 most positively and negatively differentially expressed genes by log2 fold change (p-value threshold = 5 × 10⁻⁴; fold change threshold = 4; n = 5 paired biological replicates, representative of one experiment for ( A – C ). C Normalized EGFR counts from RNA-seq data in vehicle vs agonist-treated cells. Significance determined for ( B , C ) by DESeq2 DGE analysis (Wald test (two-tailed) with Benjamini–Hochberg correction); ***p < 2×10 −52 . (Bar height = mean, error bars = SD). D Representative flow plots of EGFR expression on ILC2s with and without FFAR3 agonist. Secondary-only control: no α-EGFR primary antibody. E EGFR expression by median fluorescence intensity (MFI) (n = 5 paired biological replicates; representative of one experiment) **** p < 0.0001. F Total ILC2s after 6-day culture with DMSO AR420626 (10 µM) and gefitinib (5 µM), groups = A–C left to right. (A-B *** p = 0.0005) (B-C ** p = 0.0018). G IL-10 levels in the supernatant; two vehicle ( AR420626 -/gefitinib-) samples were below detection. (A-B **** p < 0.0001) (A–C ** p = 0.0070) (B-C ** p = 0.0097). ( F , G ): n = 5 paired biological replicates; representative of two independent experiments). H Representative intracellular cytokine staining of IL-5 (gefitinib = 10 µM). I Quantification of IL-5 MFI (n = 4 for Vehicle and AR420626 (1 sample each used as FMOs); n = 5 for AR420626 + gefitinib). IL-5 quantification for vehicle and AR420626 is shown in Fig. (A-B *** p = 0.0007) (A–C *** p = 0.0002) (B-C **** p < 0.0001). J Amphiregulin in the supernatant of ILC2s with and without 10 µM AR420626 measured by ELISA (n = 5 paired biological replicates; representative of one experiment) ** p = 0.0012. K IL-10 in supernatant of ILC2s cultured with IL-33, TSLP, IL-2 + AR420626 + α-Amphiregulin antibody or isotype control (n = 5 paired biological replicates; representative of one experiment) * p = 0.033. C , E – G , I , J bar height = mean, error bars = SD). Significance for ( E ), ( J ), and ( K ) was assessed by two-tailed paired Student’s t-test; for ( F ), ( G ), and ( I ) by two-way ANOVA with Tukey’s multiple comparisons test. ns not significant.

Article Snippet: Quantitative real-time PCR was carried out with TaqMan Gene Expression assays: Gapdh (Mm99999915) and Ffar3 (Mm02621638_s1) on a QuantStudio 5 Real-Time PCR System (Applied Biosystems) instrument for 45 cycles.

Techniques: RNA Sequencing, Two Tailed Test, Expressing, Control, Fluorescence, Staining, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Frontiers in Nutrition

Article Title: Christensenella intestinihominis MNO-863 improve obesity and related metabolic disorders via SCFAs-induced GLP-1 hormone secretion

doi: 10.3389/fnut.2025.1668786

Figure Lengend Snippet: MNO-863 regulated metabolites of cecal contents in HFD-induced obese mice. (A) 2D score plot of OPLS-DA analysis of cecum content metabolites. (B) Relative abundance of each metabolite classes in different groups in cecum content metabolomics analysis. (C) LDA score of differential cecum content metabolites by lefse analysis. (D) Heatmap analysis of significantly differential cecum content metabolites. (E) Content of the main metabolites significantly up-regulated by MNO-863 ( n = 8/group). (F) Bubble plots of MNO-863 enrichment pathway generated by MetaboAnalyst. (G,H) Representative immunohistochemistry images and quantitative analysis (Mean density) of GPR41 and GPR43 in ileum. (I) Relative expression of colonic ZO-1 and Muc-2. Data are presented as median (E) and mean ± SD (H,I) and Statistical analysis was performed using two-tail unpaired Student’s t test (E,H,I) . ns: not significant, not showed; * p < 0.05, ** p <0.01, *** p < 0.001,**** p < 0.0001.

Article Snippet: The ileum sections were individually incubated with specific antibodies against GPR41 (Proteintech, 66,811-1-IG) or GPR43 (Proteintech, 19,952-1-AQ) and HRP-conjugated secondary antibodies (DAKO, K5007).

Techniques: Generated, Immunohistochemistry, Expressing