Journal: Oncogene

Article Title: CREPT facilitates colorectal cancer growth through inducing Wnt/β-catenin pathway by enhancing p300-mediated β-catenin acetylation

doi: 10.1038/s41388-018-0161-z

Figure Lengend Snippet: CREPT enhanced CRC metastasis in vitro and in vivo. Ectopic expression of CREPT-promoted cell invasion a and migration b in HCT116 cells. Knockdown of CREPT suppressed cell invasion c and migration d in SW480 cells. e Ectopic expression of CREPT accelerated lung metastasis of HCT116 cells injected via tail vein. Pictures on the left showed representative lung morphology. HE staining showed representative lungs section. Histograph showed statistic result of metastasis nodules in nude mice lung tissues. n = 5, * P < 0.05, ** P < 0.01

Article Snippet: Human CRC cell lines (CaCO2, Colo205, DLD-1, HCT116, HT29, LoVo, SW1116, SW480, and SW620) and one immortalized colorectal epithelial cell line (NCM460) were from the American Type Culture Collection (Manassas, VA) and were cultured as described in previous study [ ].

Techniques: In Vitro, In Vivo, Expressing, Migration, Knockdown, Injection, Staining

Journal: Oncogene

Article Title: CREPT facilitates colorectal cancer growth through inducing Wnt/β-catenin pathway by enhancing p300-mediated β-catenin acetylation

doi: 10.1038/s41388-018-0161-z

Figure Lengend Snippet: CREPT regulated Wnt/β-catenin signaling pathway in CRC cells. a KEGG analysis based on RNA-sequencing data of HCT116 cells stably overexpressing CREPT or control cells. b Ectopic expression of CREPT in DLD-1 cell line enhanced TopFlash luciferase reporter activity but not FopFlash reporter ( b1 ), while depletion of CREPT reduced dual-luciferase reporter assays in SW480 cells ( b2 ). TopFlash, a classic Wnt-response luciferase reporter containing 3xTCF4 binding site. FopFlash, a negative reporter of which TCF4 binding site is mutant. c The mRNA expression of Wnt downstream targets CCND1 , c-MYC , and AXIN2 were enhanced in the presence of CREPT in DLD-1 cells and HCT116 cells. d The mRNA expression of CCND1 , c-MYC , and AXIN2 was diminished under CREPT knockdown in HT29 and SW480 cells. e The protein expression of β-catenin, c-MYC, CYCLIN D1, and AXIN2 was increased upon ectopic expression of CREPT in DLD1, HCT116 ( e1 ), HCEC 1CT, and 2CT cells ( e2 ), while decreased in CREPT knockdown HT29 and SW480 cells ( e3 ).

Article Snippet: Human CRC cell lines (CaCO2, Colo205, DLD-1, HCT116, HT29, LoVo, SW1116, SW480, and SW620) and one immortalized colorectal epithelial cell line (NCM460) were from the American Type Culture Collection (Manassas, VA) and were cultured as described in previous study [ ].

Techniques: RNA Sequencing, Stable Transfection, Control, Expressing, Luciferase, Activity Assay, Binding Assay, Mutagenesis, Knockdown

Journal: Scientific reports

Article Title: Intestinal IL-25 prevents high-fat diet-induced obesity by modulating the cholesterol transporter NPC1L1 expression in the intestinal epithelial cells.

doi: 10.1038/s41598-025-95516-7

Figure Lengend Snippet: Fig. 3. The association between intestinal IL-25 expression and the systemic and intestinal metabolic changes in high-fat diet-induced obesity. BALB/c mice were fed with NCD or HFD for 8 weeks. The alteration of intestinal cytokine gene expression was determined and associated with various metabolic parameters. (A) Quantitative real-time PCR analysis of proinflammatory cytokine (Tnfa) and epithelial cell- derived cytokines (Il25, Tslp, Il33) in the jejunum of NCD-and HFD-fed mice. The mRNA expression data are presented as fold induction over actin (Actb) expression, with the mRNA levels in NCD-fed mice set as 1. Graphs depict mean ± SD of three independent experiments, with n = 6 mice per group. Significance was determined using Student’s t-test analysis. (B) The correlation between plasma glucose, total cholesterol (TC), and triglycerides (mg/dL) and the mRNA expression levels of intestinal cytokines Tnfa (upper row) and Il25 (bottom row), with n = 10 mice from HFD group was analyzed using Spearman’s rank test. (C) The association between the expression level of Il25 gene and genes related to glucose and lipid absorption in the jejunum, including Sglt1, Npc1l1, Fatp4, Cd36, with n = 10 mice from HFD group was analyzed using Spearman’s rank test. Correlation coefficients (r) and p values are provided. (*p < 0.05, **p < 0.01)

Article Snippet: Human epithelial cell lines (Caco-2 cells, ATCC HTB-37) were grown in high-glucose Dulbecco’s Modified Eagle Medium (DMEM, GibcoTM), supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% penicillin-streptomycin (P/S).

Techniques: Expressing, Gene Expression, Real-time Polymerase Chain Reaction, Derivative Assay, Clinical Proteomics

Journal: Mucosal Immunology

Article Title: Fundamental role for HIF-1α in constitutive expression of human β defensin-1

doi: 10.1038/mi.2013.6

Figure Lengend Snippet: Physiological hypoxia and basal hypoxia-inducible factor (HIF) maintain mouse and human defensin expression. ( a ) Pimonidazole HCl forms adducts in tissue experiencing tissue pO 2 ⩽10 mm Hg and enabled immunohistochemical visualization (red) of “physiological hypoxia” in brush-border epithelia lining the colon of healthy C57BL/6J mice. Bar=100 μ M . ( b ) This physiological hypoxia maintains expression of murine defensin genes as evidenced by reduced expression in epithelial scrapings from conditional Hif1a −/− mice vs. control. ( c ) Human intestinal epithelial cell lysate (Caco-2) contained detectable HIF-1α, even in normoxic conditions, which was increased by overnight incubation in hypoxia. ( d ) A screen of defensin expression in Caco-2 cells with lentiviral ARNT (HIF-1β) knock down (KD) revealed a role for basal HIF in beta-defensin 1 ( DEFB1 ) expression. Panel b performed with three animals per group. Panel d represents five independent experiments. Error bars represent s.e.m. * P <0.05, ** P <0.01, *** P <0.001 as determined by one-way analysis of variance ( b ) and paired Student's t -test ( d ). Sh, short hairpin.

Article Snippet: Caco-2 (ATCC# HTB-37) and T84 (ATCC# CCL-248) human intestinal epithelial cells were obtained from ATCC (Manassas, VA) and maintained in 95% air with 5% CO 2 at 37 °C according to ATCC's instructions.

Techniques: Expressing, Immunohistochemical staining, Control, Incubation, Knockdown

Journal: Mucosal Immunology

Article Title: Fundamental role for HIF-1α in constitutive expression of human β defensin-1

doi: 10.1038/mi.2013.6

Figure Lengend Snippet: Hypoxia-inducible factor-1α (HIF-1α) is critical for beta-defensin 1 ( DEFB1 ) expression. Lentiviral HIF-1α and HIF-2α knock down (KD) of Caco-2 (previously validated) and T84 cells (knock down shown in ( a )) reveal that loss of HIF-1α, but not HIF-2α, reduced DEFB1 expression in ( b ) Caco-2 and ( c ) T84 intestinal epithelial cells. Exposure to hypoxia (1% O 2 ) did not induce DEFB1 . ( d ) Co-transfection of HIF-1α or HIF-2α plasmids containing mutated oxygen-dependent degradation domains with a HRE (hypoxia response element) luciferase reporter plasmid demonstrates the ability of oxygen-stable HIF to stimulate promoter activity. However, ( e ) HIF plasmid co-transfection did not stimulate DEFB1 promoter activity, possibly due to basal HIF saturation. ( f ) Ascorbate supplementation (1 mM, 24 h) did not influence cytoplasmic localization of NF-κB (p65). Shown here are three independent samples with (+) and without (−) ascorbate. Regardless, supplementation with the prolyl-hydroxylase cofactor ascorbate (Asc; 1 m M for 24 h) reduced ( g ) cytoplasmic and nuclear HIF-1α and ( h ) DEFB1 expression consistent with the model that basal HIF saturates the DEFB1 promoter. All figures ( a – h ) represent the mean of at least three independent experiments. Error bars represent s.e.m. * P <0.05, *** P <0.0001 as determined by one-way analysis of variance comparing HIF-1α knockdown to baseline short hairpin (Sh) control ( b , c ) and paired Student's t -test ( c , d ). NF, nuclear factor.

Article Snippet: Caco-2 (ATCC# HTB-37) and T84 (ATCC# CCL-248) human intestinal epithelial cells were obtained from ATCC (Manassas, VA) and maintained in 95% air with 5% CO 2 at 37 °C according to ATCC's instructions.

Techniques: Expressing, Knockdown, Cotransfection, Luciferase, Plasmid Preparation, Activity Assay, Control

Journal: Mucosal Immunology

Article Title: Fundamental role for HIF-1α in constitutive expression of human β defensin-1

doi: 10.1038/mi.2013.6

Figure Lengend Snippet: Impact of hypoxia-inducible factor-1α ( HIF1a ) knock down (KD) on functional human β defensin-1 (hBD-1) protein. ( a ) HIF-1α knock down reduced hBD-1 staining in both Caco-2 and T84 intestinal epithelial cell lines compared with short hairpin (Sh) control or HIF-2α knockdown. ( b ) HIF-1α knock down also decreased hBD-1 secreted into cell supernatant as measured by enzyme-linked immunosorbent assay. ( c ) A bacterial killing assay, using Escherichia coli Nissle, which is sensitive to oxidized recombinant hBD-1 and resistant to hyposmotic stress from dilution of salt that inactivates hBD-1, revealed a relevance of a 50% reduction in recombinant hBD-1 on bacterial viability. All figures ( a – c ) are representative ( a ) or the mean ( b , c ) of at least three independent experiments. Error bars represent s.e.m. ** P <0.01 as determined by one-way analysis of variance ( b ). CFU, colony-forming unit; IgG, immunoglobulin G.

Article Snippet: Caco-2 (ATCC# HTB-37) and T84 (ATCC# CCL-248) human intestinal epithelial cells were obtained from ATCC (Manassas, VA) and maintained in 95% air with 5% CO 2 at 37 °C according to ATCC's instructions.

Techniques: Knockdown, Functional Assay, Staining, Control, Enzyme-linked Immunosorbent Assay, Recombinant