Journal: Aging and Disease

Article Title: Human Adipose Mesenchymal Stem Cell-derived Exosomes Protect Mice from DSS-Induced Inflammatory Bowel Disease by Promoting Intestinal-stem-cell and Epithelial Regeneration

doi: 10.14336/AD.2021.0601

Figure Lengend Snippet: hADSC-Exo reshape the cell structure in colon crypts of DSS-induced mice IBD . ( A-D ) Immunofluorescence staining analysis of expression of epithelial cells proliferation related specific marker Ki67 and β-Catenin in mice colon tissue sections (scale bar=20 µm; n=7, per group). ( E-H ) Immuno-fluorescence staining analysis of expression of colon stem cells specific markers EPH B2 and LGR5 in mice colon tissue sections (scale bar=20 µm; n=7, per group). (I and J) Immuno-fluorescence staining analysis of expression of goblet cells specific marker Mucin in mice colon tissue sections (scale bar=20 µm; n=7, per group).

Article Snippet: Hcoepic epithelial cell proliferation assay was performed by cell counting kit-8 (CCK8, Dojindo Molecular Technologies) following manufacture instructions.

Techniques: Immunofluorescence, Staining, Expressing, Marker, Fluorescence

Journal: Aging and Disease

Article Title: Human Adipose Mesenchymal Stem Cell-derived Exosomes Protect Mice from DSS-Induced Inflammatory Bowel Disease by Promoting Intestinal-stem-cell and Epithelial Regeneration

doi: 10.14336/AD.2021.0601

Figure Lengend Snippet: hADSC-Exo up-taken by Hcoepic cells and promotes wound healing of Hcoepic cells . ( A ) Uptake of hADSC-Exo by Hcoepic cells at 4, 12, 24, 48 h after co-culture. ( B ) Mean fluorescence intensity (MFI) of Dil in Hcoepic cells. (C and D) hADSC-Exo promotes the migratory activity of Hcoepic cells and accelerate wound healing, at 0, 4, 6, 12, 24, 48 h after co-culture.

Article Snippet: Hcoepic epithelial cell proliferation assay was performed by cell counting kit-8 (CCK8, Dojindo Molecular Technologies) following manufacture instructions.

Techniques: Co-Culture Assay, Fluorescence, Activity Assay

Journal: Aging and Disease

Article Title: Human Adipose Mesenchymal Stem Cell-derived Exosomes Protect Mice from DSS-Induced Inflammatory Bowel Disease by Promoting Intestinal-stem-cell and Epithelial Regeneration

doi: 10.14336/AD.2021.0601

Figure Lengend Snippet: hADSC-Exo facilitates Hcoepic cells proliferation, promotes cell cycle and inhibits cell apoptosis . (A and B) CCK8 assay analysis the exosomes’ influences on the proliferation of Hcoepic cells at 24 (A) & 48 h (B). exosome concentration: 0, 0.1, 2, 5, 8, 10 µg/ml. ( C ) Exosomes’ influences on cell cycle of Hcoepic cells after hADSC-Exo administration 48 h at exosome concentration: 200 ug/ml. ( D ) Exosomes’ influences on cell apoptosis of Hcoepic cells after hADSC-Exo administration 48 h at exosome concentration: 200 µg/ml (All experiments repeated at least 3 times).

Article Snippet: Hcoepic epithelial cell proliferation assay was performed by cell counting kit-8 (CCK8, Dojindo Molecular Technologies) following manufacture instructions.

Techniques: CCK-8 Assay, Concentration Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA Loc554202 induces apoptosis in colorectal cancer cells via the caspase cleavage cascades

doi: 10.1186/s13046-015-0217-7

Figure Lengend Snippet: Relative expression of Loc554202 in colorectal cancer tissues and cells compared with adjacent normal tissues and normal colonic epithelial cells. a The relative expression of Loc554202 in colorectal cancer tissues ( n = 48) compared with corresponding non-tumor tissues ( n = 48). The Loc554202 expression was examined by qRT-PCR and was normalized to the GAPDH expression. The results are presented as the fold-change in tumor tissues relative to normal tissues (shown as -ΔΔCT). b and c ) The data are presented as the relative expression levels in tumor tissues. The Loc554202 expression was significantly lower in patients with a higher pathological stage and larger tumors. d The Loc554202 expression was assessed by qRT-PCR in colorectal cancer cell lines (HT-29, RKO, SW480, HCT116, DLD1) and was compared with the normal human colonic epithelial cell line (HCoEpiC). e The relative expression levels of Loc554202 following the treatment of HCT116 and DLD1 cells with pCDNA-Loc554202 and empty vector. ( f and g ) The level of Loc554202 expression in HCT116 and DLD1 cells following treatment with 5-aza-dC (0, 5, 10 μM).* P < 0.05 and ** P < 0.01

Article Snippet: The human colonic epithelial cells HCoEpiC were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Quantitative RT-PCR, Plasmid Preparation

Journal: Cell Death & Disease

Article Title: Ferroptosis involves in intestinal epithelial cell death in ulcerative colitis

doi: 10.1038/s41419-020-2299-1

Figure Lengend Snippet: a Cell death of HCoEpiC cells with or without RSL3 (20 μ m ) challenged for 8 hours was measured by PI staining, positive signal was stained in red (Scale: 50 μm). b ROS of HCoEpiC cells with or without RSL3 treatment was detected by C11-BODIPY staining, positive signal was stained in red (Scale: 50 μm). c Western blotting analysis of p-eIF2α, eIF2α, FTL, and FTH in cells with RSL3 challenged in the indicated times. β-actin was used as the loading control. d Relative mRNA level of GPR78, ATF4, CHOP, PTGS2, FTL, and FTH in HCoEpiC cells were measured by real-time PCR. Cells were treated with or without RSL3 for 8 h, GSK2606414 (GSK 414, 1 μ m ) was added to the cells 30 mins before RSL3. e Western blotting analysis of p-eIF2α, eIF2α, FTL, and FTH in cells with or without RSL3 and GSK 414 treatment. β-actin was used as the loading control. f Cell death of indicated cells was tested through PI staining (Scale: 50 μm). g ROS of indicated cells was detected by C11-BODIPY staining (Scale: 50 μm). Statistical analyses were performed with Student’s t tests (two groups) or one-way ANOVA (more than two groups).

Article Snippet: The HCoEpiC cell (human normal colonic epithelial cell) was cultured in colonic epithelial cell medium (CoEpiCM, ScienCell Research Laboratory, CA, USA) containing 10% fetal bovine serum and other supplements according to the manufacturer's instructions (ScienCell Research Laboratory).

Techniques: Staining, Western Blot, Control, Real-time Polymerase Chain Reaction

Journal: Cell Death & Disease

Article Title: Ferroptosis involves in intestinal epithelial cell death in ulcerative colitis

doi: 10.1038/s41419-020-2299-1

Figure Lengend Snippet: a Necrotic cell death in colonic epithelial tissues from p65 IEC-KO or WT mice was labeled by PI and analyzed through flow cytometry. b , c MDA levels and iron levels were detected in colonic epithelial tissues from p65 IEC-KO or WT mice received DSS administration. d , e Western blotting analysis of p65, p-eIF2α, eIF2α, FTL, and FTH in colonic epithelial/non-epithelial tissues from p65 IEC KO or WT mice after DSS treatment. β-actin was used as the loading control. f Double immunofluorescence staining for p-eIF2α and CK 18 in colonic sections of p65 IEC KO or WT mice with DSS challenged. Nuclei was stained with DAPI in blue. p-eIF2α was stained in green and CK 18 was visualized in red, the coincident positive signals were visualized in yellow (Scale: 50 μm). g Double immunofluorescence staining for FTH and CK 18 in colonic sections. FTH was stained in green, and CK 18 was visualized in red, the merging positive signals were visualized in yellow (Scale: 50 μm). h Double immunofluorescence staining for p-eIF2α and FTH in colonic sections. p-eIF2α was stained in green and FTH was visualized in red, the coincident positive signals were visualized in yellow (Scale: 50 μm). i Western blotting of p-p65, p65 p-eIF2α, and eIF2α in HCoEpiC cells with or without TNF-α treatment (40 ng/ml, 1 h). β-actin was used as the loading control. j Immunofluorescence staining for p-p65 and p-eIF2α in HCoEpiC cells. Nuclei was stained with DAPI in blue. p-p65 was stained in green and p-eIF2α was visualized in red, the overlapped positive signals were visualized in yellow (Scale: 50 μm). k HCoEpiC cells with or without TNF-α challenged were subjected to immunoprecipitation with an anti-p65 antibody. Co-immunoprecipitated endogenous p-p65, eIF2α, and p-eIF2α were detected with corresponding antibodies. IP immunoprecipitate, IB immunoblot. l Analysis of the effect of BAY 11-7085 (10 μ m , adding to the indicated wells 15 mins before TNF-α administration) on the activation of p65 and eIF2α in cells with or without TNF-α treatment through western blotting. β-actin was used as the loading control. Statistical analyses were performed with Student’s t tests (two groups) or one-way ANOVA (more than two groups).

Article Snippet: The HCoEpiC cell (human normal colonic epithelial cell) was cultured in colonic epithelial cell medium (CoEpiCM, ScienCell Research Laboratory, CA, USA) containing 10% fetal bovine serum and other supplements according to the manufacturer's instructions (ScienCell Research Laboratory).

Techniques: Labeling, Flow Cytometry, Western Blot, Control, Double Immunofluorescence Staining, Staining, Immunofluorescence, Immunoprecipitation, Activation Assay