human intestinal epithelial cells  (ATCC)

Journal: mBio

Article Title: Intestinal epithelial Tet2 deficiency reprograms the gut microbiota through bile acid metabolic alterations

doi: 10.1128/mbio.03562-25

Figure Lengend Snippet: The impact of intestinal epithelial Tet2 deficiency on epithelial architecture, bile acid metabolism, and gut microbiota composition. ( A ) Representative hematoxylin and eosin (H&E) stained images of the ileum and colon are presented. Tet2-iKO mice exhibit submucosal gaps at the villus tips (indicated by red arrows) and focal epithelial shedding with exposed lamina propria in the colon (indicated by black arrows) compared to wild-type (WT) mice. ( B and C ) Quantitative analysis of crypt depth in the ileum ( B ) and muscularis thickness in the colon ( C ) is provided ( n = 4). ( D ) The relative expression levels of epithelial lineage marker genes (Lgr5, Alpi, Clca1, Muc2, Chga, Vil1) in the ileum are shown ( n = 5). ( E ) There is a downregulation of bile acid transporter genes (Slc10a2) in Tet2-iKO mice ( n = 5). ( F ) In Caco-2 cells, Tet2 knockdown or inhibition using the Tet2 inhibitor Bobcat 339 results in decreased ASBT protein levels, whereas Tet2 activation via ascorbic acid leads to increased ASBT expression. ( G ) Dot blot analysis of 5hmC levels in control versus Tet2-knockdown Caco-2 cells ( n = 3). ( H ) Chromatin immunoprecipitation (ChIP)-qPCR showing Tet2 enrichment at the Slc10a2 promoter region in Caco-2 cells; IgG was used as a control ( n = 3). ( I ) Young Tet2-iKO mice exhibit increased fecal bile acid excretion and elevated bile acid levels in plasma and liver compared to WT littermates ( n = 5). ( J ) Hepatic expression of the bile acid synthesis enzyme Cyp27a1 ( n = 4). ( K ) Metabolomic profiling reveals elevated fecal hyocholic acid (HCA) in Tet2-iKO mice. ( L ) Molecular docking predicts favorable binding of HCA within the ASBT binding site. ( M ) Surface plasmon resonance (SPR) confirms direct binding between HCA and ASBT. ( N and O ) 16S rRNA sequencing shows altered microbiota composition and increased abundance of Lactobacillus in Tet2-iKO mice ( n = 3). ( P ) In vitro bacterial growth assay demonstrates the dose-dependent effect of HCA on Lactobacillus proliferation. Differences between the two groups were assessed by Student’s t -test; multiple groups were compared by one-way ANOVA. Alpha-diversity indices (ACE, Chao1, Simpson) were analyzed using the Wilcoxon rank-sum test. Data are mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human intestinal epithelial Caco-2 cells (ATCC HTB-37) were maintained in DMEM (Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco), 1% non-essential amino acids, and 1% penicillin-streptomycin at 37°C in 5% CO 2 .

Techniques: Staining, Expressing, Marker, Knockdown, Inhibition, Activation Assay, Dot Blot, Control, Chromatin Immunoprecipitation, ChIP-qPCR, Clinical Proteomics, Metabolomic, Binding Assay, SPR Assay, Sequencing, In Vitro, Growth Assay

Journal: Scientific Reports

Article Title: Bee pollen-derived peptide with dual DPP-IV Inhibition and glucose transport modulation

doi: 10.1038/s41598-026-39009-1

Figure Lengend Snippet: The percentage viability of Caco-2 cells follow 48 h exposure to AA-7 ( a ) and glucose uptake inhibition kinetics with fluorescent glucose 2-NBDG, following treatment with AA-7 ( b ). Data are presented as mean ± SD for three or more independent trials with consistent findings. The use of different letters (a-c) identifies means which exhibit significant differences ( p < 0.05).

Article Snippet: Human intestinal epithelial cell line Caco-2 cells (ATCC ® HTB-37TM) were supplied by the American Type Culture Collection (ATCC, Manassas, VA, USA), before being cultured in Eagle’s Minimum Essential Medium (EMEM) which was supplemented using L-glutamine and 10% fetal bovine serum (Gibco, Rockville, MD, USA).

Techniques: Inhibition