Journal: Food Science & Nutrition

Article Title: Protective Effects of Nongxiang‐Type Tieguanyin on the Liver of D‐Galactose‐Induced Aging C57BL /6 Mice via the NF ‐ κB Signaling Pathway

doi: 10.1002/fsn3.72396

Figure Lengend Snippet: Tgy‐N reduces cellular senescence markers in Nutlin‐3a‐induced senescent AML12 and HepG2 cells. (A) SA‐β‐gal staining of AML12 cells. Scale bar = 100 μm. (B) Quantitative analysis of SA‐β‐gal‐positive AML12 cells. (C) Representative Western blots of p16, p21, and GAPDH in AML12 cells. (D) Quantitative analysis of p16/GAPDH ratio in AML12 cells. (E) Quantitative analysis of p21/GAPDH ratio in AML12 cells. (F) SA‐β‐gal staining of HepG2 cells. Scale bar = 100 μm. (G) Quantitative analysis of SA‐β‐gal‐positive HepG2 cells. (H) Representative Western blots of p16, p21, and GAPDH in HepG2 cells. (I) Quantitative analysis of p16/GAPDH ratio in HepG2 cells. (J) Quantitative analysis of p21/GAPDH ratio in HepG2 cells. Data are expressed as mean ± SEM. Statistical assessments were performed using one‐way ANOVA followed by post hoc Duncan's test. Different letters above the bars denote statistically significant differences between groups at the p < 0.05 level.

Article Snippet: AML12 (mouse normal hepatocytes) and HepG2 (human hepatocellular carcinoma) cell lines were obtained from ATCC.

Techniques: Staining, Analysis, Western Blot

Journal: Nature Communications

Article Title: Trifluoroacetate mediated reduction of atherosclerosis in mice

doi: 10.1038/s41467-026-77341-2

Figure Lengend Snippet: a Expression of several PPAR target genes in rat FAO liver cells was increased by TFA treatment, as indicated by qPCR. The effect was lost in PPAR-α knockout FAO cells. The data are shown as mean ± SD for technical triplicates. p values were determined by one-way ANOVA comparing each experimental group to the no treatment control, post hoc Dunnett’s test. The experiment was repeated twice independently with similar results. b Reporter assay in human HepG2 liver cells transfected with different mouse PPAR isoforms, in which luciferase expression is driven by activation of the PPAR response element. A different positive control was used for each PPAR isoform: 0.1 mM ciprofibrate for PPAR-α, 1 µM GW501516 for PPAR-δ/β, and 10 µM rosiglitazone for PPAR-γ. The data are shown as mean ± SD for technical triplicates. p values were determined by one-way ANOVA comparing each experimental group to the no treatment control, post hoc Dunnett’s test. The experiment was repeated twice independently with similar results. c A coactivator recruitment assay based on biolayer interferometry (BLI) was used to determine if TFA promotes binding of purified PPAR-α to a coactivator peptide. On the left are shown BLI association/dissociation traces for different putative ligands of PPAR-α. On the right is a plot of binding responses (maximum shift in the association phase) for palmitic acid and TFA, which had EC 50 values of 0.2 and of 4.9 mM, respectively. Acetate did not promote binding at the highest concentration tested of 10 mM. Data are shown as mean ± SD for technical triplicates. The experiment was repeated twice independently with similar results. d Data including RNA-seq, confocal microscopy, western blotting, metabolomics, BLI binding assays, and biochemical assays indicated that TFA induces peroxisome proliferation by activating peroxisome proliferator-activated receptor (PPAR)-α. e While TFA is structurally similar to acetate, we did not observe trifluoroacetylated metabolites or proteins, nor trifluoroacetyl-CoA, using a combination of metabolomics, proteomics, and biochemical assays. f TFA treatment did not alter the gut microbiome composition, as determined by 16S sequencing of fecal microbiome samples from PBS vehicle- or TFA-treated wild-type C57BL/6J mice (4-week treatment, 27 mg/kg/day TFA, n = 10–15/group). On the left is shown a Bray-Curtis principal component analysis of beta diversity between samples. On the right, plots of Chao1 index alpha diversity are shown as mean ± SD. Each datapoint represents one biological replicate; sample sizes were CHOW/PBS vehicle ( n = 10), HFD/PBS vehicle ( n = 15), HFD/TFA ( n = 10). p values were determined by one-way ANOVA comparing the experimental group to PBS vehicle group, post hoc Sidak’s multiple comparison test; ns not significantly different.

Article Snippet: The HepG2 cell line (obtained from ATCC, cat. # HB-8065, RRID: CVCL_0027) was cultured in DMEM medium (Gibco) containing 10% FBS (HyClone), 100 U/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Expressing, Knock-Out, Control, Reporter Assay, Transfection, Luciferase, Activation Assay, Positive Control, Binding Assay, Purification, Concentration Assay, RNA Sequencing, Confocal, Microscopy, Western Blot, Microbiome, Sequencing, Analysis, Comparison