Journal: Chemmedchem

Article Title: Scaffold Hopping Provides Liver X Receptor Agonist With Preference for the β Subtype

doi: 10.1002/cmdc.70509

Figure Lengend Snippet: In vitro characterization of the optimized LXR agonist 7 . (a) Full‐length LXR reporter gene assay in HEK293T cells. N ≥ 3. (b) Quantification of LXR target genes in HepG2 cells. Both 2 and ( S )‐ 7 robustly induced expression of target genes involved in cholesterol metabolism (ABCA1, ABCG1) and DNL (FAS, SREBP1c). Data are the mean ± SEM of N = 4; ** p < 0.01, *** p < 0.001 (ANOVA with post hoc Bonferroni test against 0.1% DMSO or as indicated by the bars). Full agonist T0901317 ( 32 ) was tested for comparison. (c,d) Lipogenesis assay. Incubation of HepG2 cells with pan LXR full agonist T0901317 ( 32 ) for 72 h led to a substantial accumulation of lipids that were stained with Oil Red O. In comparison to 32 , both the parent compound 2 and ( S )‐ 7 displayed less lipogenic potential. Data are the mean ± SEM of N = 3; * p < 0.05, ** p < 0.01, *** p < 0.001 (one‐sided t ‐test against 0.1% DMSO). The images shown in (d) are representative details from the full images that can be found in Figure S3. (e) In a cytotoxicity assay, 2 and ( rac )‐ 7 did not influence the metabolic activity of HepG2 cells but induced cytotoxicity to HEK293T cells in the micromolar concentration region. While these concentrations are well above the EC 50 value of ( S )‐ 7 in the Gal4 reporter gene assay (red dashed line), compound 2 showed partially toxic effects at its EC 50 value (green dashed line). (f) Metabolic stability of 2 and ( rac )‐ 7 in rat liver microsomes. After 60 min, ( rac )‐ 7 was almost recovered completely, whereas approximately half of 2 had been metabolized. 7‐Ethoxycoumarine (7‐EC) served as internal standard. Data are the mean ± SEM of N = 3.

Article Snippet: HEK293T and HepG2 cells (German Collection of Microorganisms and Cell Cultures, DSMZ) were grown in Dulbecco's modified Eagle's medium (DMEM) high glucose supplemented with 10% fetal bovine serum (FBS), sodium pyruvate (1 mM), penicillin (100 U/mL), and streptomycin (100 μg/mL) at 37 °C and 5% CO 2 .

Techniques: In Vitro, Reporter Gene Assay, Expressing, Comparison, Incubation, Staining, Cytotoxicity Assay, Activity Assay, Concentration Assay, Stability

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