Journal: Chinese Medicine

Article Title: Sanren decoction ameliorates ulcerative colitis by modulating gut microbiota and macrophage polarization to enhance intestinal barrier function

doi: 10.1186/s13020-025-01183-1

Figure Lengend Snippet: FMT enhances intestinal barrier integrity and facilitates the polarization of M2 macrophages. A – E Expression of Claudin-5, Occludin, ZO-1, and PTGS2 proteins in mouse colonic tissue by WB (n = 3). F Expression of Muc2 protein by immunohistochemistry. G – I Expression of iNOS and ARG1proteins in mouse colonic tissue by WB (n = 3). J Immunofluorescence staining of M1 macrophages in mouse colonic sections. K Immunofluorescence staining of M2 macrophages in mouse colonic sections. Statistical significance indicated by * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: IL-6 Detection Kit (Nanjing Bioyntech, BY- EM220188 ); TNF-α Detection Kit (Nanjing Bioyntech, BY- EM220852 ); IL-1β Detection Kit (Nanjing Bioyntech, BY-EM2201740); β-Actin Monoclonal Antibody (Proteintech, 66009-1-Ig, 1:2000); Occludin Polyclonal Antibody (Proteintech, 27260-1-AP, 1:2000); ZO-1 Polyclonal Antibody (Proteintech, 21773-1-AP, 1:2000); Claudin-5 Polyclonal Antibody (Proteintech, 29767-1-AP, 1:2000); Anti-COX2/Cyclooxygenase 2 Rabbit mAb (PTMBIO, PTM-7147, 1:1000); Anti-iNOS antibody[RM1017] (abcam, ab283655, 1:2000); Arginase-1 Polyclonal antibody(Proteintech,1600–1-AP, 1:5000); GAPDH Rabbit mAb (zenbio, R380626, 1:5000); Anti-beta Tubulin Rabbit pAb (Servicebio,GB11017,1:2000); CD86 (SAB, 48763-1, 1:400); CD163 (Proteintech, 16646-1-AP, 1:400); F4/80 (Zenbio, 263101, 1:400); MUC2 Ready-to-use Reagent Kit for IHC (Proteintech, KHC1327, 1:1200); Goat Anti-Mouse IgG H&L (UE, H6161, 1:8000); Goat Anti-Rabbit IgG H&L (UE, H6161, 1:8000); Two-Step Immunohistochemistry Detection Kit (BOSTER, SV0002); DyLight 594 Conjugated AffiniPure Goat Anti-Mouse IgG H&L (BOSTER, BA1141, 1:400); iFluorTM 488 Conjugated Goat Anti-Rabbit IgG Polyclonal Antibody (HUABIO, HA1121, 1:400); Ampicillin (TargetMol, T0814L); Vancomycin (TargetMol, T8641); Neomycin Sulfate (TargetMol, T0950); Metronidazole (TargetMol, T1079);DSS for dextran sulfate sodium salt colitis modeling (meilunbio, MB5535-2);16S rRNA and untargeted metabolomics sequencing was provided by Metware Biotechnology Co., Ltd. (Wuhan, China); Mice were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd. (Nanjing, China); The mice were housed at the Laboratory Animal Science Center of Nanchang University (Nanchang, China).

Techniques: Expressing, Immunohistochemistry, Immunofluorescence, Staining

Journal: Molecules

Article Title: A Novel ERK2 Degrader Z734 Induces Apoptosis of MCF–7 Cells via the HERC3/p53 Signaling Pathway

doi: 10.3390/molecules27144337

Figure Lengend Snippet: Z734 causes apoptosis and inhibits MCF–7 cells proliferation and migration. ( A ) The chemical structure of Z734. ( B ) Apoptosis levels were detected via flow cytometry. WT MCF–7 cells were treated with Z734 (0, 5, 10, and 15 μM) for 12 h. Flow cytometric analysis of apoptotic cells was conducted using AnnexinV–FITC. ( C ) Measurement of Caspase3 activity after Z734 intervention in MCF–7 cells. ( D ) Apoptosis–related protein levels after treatment with Z734 were determined via Western blot analysis. ( E ) MCF–7 cells were treated with different doses of Z734 and incubated for 12 h. Cell proliferation was assessed via CCK–8 assays. ( F ) Ki67 distribution after treatment with Z734 was determined via immunofluorescence. Quantification of fluorescence was performed on the average optical density using the ImageJ software. Scale bars = 10 μm. ( G ) The percentage of colonies of MCF–7 cell lines was determined after treatment with Z734. ( H ) MCF–7 cells were treated with Z734 for 12 h and then analyzed for migration by transwell analysis. Scale bars = 20 μm. Data shown in ( B , C , E , G , H ) were used one–way ANOVA with Tukey’s post hoc test. Data shown in ( F ) was used two–tailed Student’s t –test. All assays were performed in triplicate, and the data are expressed as mean ± standard deviation. * p < 0.05, ** p < 0.01, *** p < 0.001, n.s., no significant difference.

Article Snippet: Human Ki67 ELISA kits (ab253221) were purchased from Abcam, Wuhan, China.

Techniques: Migration, Flow Cytometry, Activity Assay, Western Blot, Incubation, CCK-8 Assay, Immunofluorescence, Fluorescence, Software, Two Tailed Test, Standard Deviation

Journal: Molecules

Article Title: A Novel ERK2 Degrader Z734 Induces Apoptosis of MCF–7 Cells via the HERC3/p53 Signaling Pathway

doi: 10.3390/molecules27144337

Figure Lengend Snippet: A combination of Z734 and lapatinib inhibits the growth of breast cancer. ( A ) MCF–7 cells were incubated with Z734 (10 μM) and/or lapatinib (0.5 μM), and p–ERK2, ERK2, p–p53, p53 and HERC3 protein expressions were detected by WB. ( B ) Representative images of ERK2 and p53 immunofluorescence staining in MCF–7 treated with Z734 (10 μM) or lapatinib (0.5 μM). DAPI was used to stain the nuclei. Quantification of fluorescence was performed on the Average Optical Density using the ImageJ software. Scale bars = 10 μm. ( C ) ELISA was used to detect the expression levels of Ki67 in MCF–7 to reflect cell proliferation after Z734 and/or lapatinib treatment. ( D ) Caspase3 activities in overexpressing of ERK2 in MCF–7 treated with Z734 (10 μM) and/or lapatinib (0.5 μM) were detected. ( E ) MCF–7 were treated with Z734 (10 μM) and/or lapatinib (0.5 μM) for 12 h, followed by flow cytometry to detect cell apoptosis. ( F ) Migration of MCF–7 cells were evaluated using the transwell assay after being treated with Z734 (10 μM) and/or lapatinib (0.5 μM). Scale bars = 20 μm. ( G ) Body weights of female Balb/c mice with different treatment groups ( n = 5 for Control group; n = 5 for Z734 group; n = 5 Lapatinib group; n = 5 for Z734 + Lapatinib group). ( H ) Picture ( n = 5 for Control group; n = 5 for Z734 group; n = 5 Lapatinib group; n = 5 for Z734 + Lapatinib group, combined results of 2 independent experiments) of MCF–7 breast tumor across four groups. ( I , J ) The volumes ( I) and weights ( J) were measured and quantified ( n = 5 for Control group; n = 5 for Z734 group; n = 5 Lapatinib group; n = 5 for Z734 + Lapatinib group). Data shown in ( B – G , I , J ) were used one–way ANOVA with Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human Ki67 ELISA kits (ab253221) were purchased from Abcam, Wuhan, China.

Techniques: Incubation, Immunofluorescence, Staining, Fluorescence, Software, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Migration, Transwell Assay

Journal: Cell Death & Disease

Article Title: Claudin-7 is essential for the maintenance of colonic stem cell homoeostasis via the modulation of Wnt/Notch signalling

doi: 10.1038/s41419-024-06658-x

Figure Lengend Snippet: A – D Normalised enrichment scores (NES) from GSEA and enrichment plots for intestinal stem cells, differentiated cells, inflammatory responses, and apoptosis transcriptional signatures of Cldn-7 fl/fl ;Villin-CreERT2 and Cldn-7 fl/fl ;Villin-CreW colonic tissues, respectively. E Heatmap showing the expression levels of established genes in Wnt signalling pathways, stem cells, and mature differentiated cells in either Cldn-7 fl/fl ;Villin-CreERT2 or Cldn-7 fl/fl ;Villin-CreW colonic tissues. F Differential expression analysis of Log 2 fold change in stem cells and Wnt signalling pathway by Cldn-7 deficiency. G Representative images of colonic organoids derived from Cldn-7 fl/fl ;Villin-CreERT2 and Cldn-7 fl/fl ;Villin-CreW mice (Left) from a minimum of three independent experiments; scale bars: 50 μm. The right panel shows the quantification of the average colonic organoid size on day 7 of the first passage. Organoids size was randomly measured in 50 organoids. H Quantitative RT-PCR analysis of marker genes for ISCs and differentiated cells in organoids derived from control mice and Cldn-7 knockout mice. ( n = 3–4 per genotype). All images are representative of at least three independent experiments. Statistical data are presented as mean ± SEM. All p values were calculated using Student’s t -test (two-tailed) or Mann–Whitney nonparametric test; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns, not significant.

Article Snippet: A terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling (TUNEL) assay was performed using a one-step TUNEL apoptosis assay kit (Beyotime), according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative Proteomics, Derivative Assay, Quantitative RT-PCR, Marker, Control, Knock-Out, Two Tailed Test, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: Claudin-7 is essential for the maintenance of colonic stem cell homoeostasis via the modulation of Wnt/Notch signalling

doi: 10.1038/s41419-024-06658-x

Figure Lengend Snippet: A Representative fluorescence images of Lgr5-EGFP and Cldn-7 staining of intestinal crypts. Arrowheads denote Cldn-7 expression (red) in Lgr5 + ISCs (EGFP); scale bars: 20 μm. B RNAscope analysis confirms the effects of Cldn-7 deletion (green) in the crypt region, detected across a minimum of three independent replicates; scale bars: 20 μm. C Immunofluorescence images of distal colonic tissues stained for Cldn-7 (red) and Lgr5-EGFP (green); cell nuclei were counterstained with DAPI (blue). The arrow indicates the absence of Cldn-7 in the crypt area; scale bars: 20 μm. Graphs (right) present the quantification of Lgr5 + ISCs in the intestinal crypts ( n = 3 per genotype). D Quantitative RT-PCR analysis of Cldn-7 in sorted Lgr5-EGFP + ISCs by flow cytometry in colonic tissues from Cldn-7 fl/fl ;Lgr5-CreERT2 mice after VEH or TAM administration. ( n = 5 per genotype). E Changes in body weight ( n = 7 per genotype) and colon length ( n = 9 per genotype) during TAM induction. F Representative images of pathological HE staining, Ki-67 staining, and TUNEL assays of colon sections from Cldn-7 fl/fl ;Lgr5-CreERT2 and Cldn-7 fl/fl ;Lgr5-CreW mice following TAM induction for 5 days (D5) and recovery for 1 week (D12). Arrowheads denote intestinal epithelial disruption. Images are representative of 3 mice per genotype; scale bars: 50 μm. Right panels: quantification of Ki-67-positive cells per crypt base and TUNEL-positive cells per field. G RNAscope in situ hybridisation of Lgr5 (red) and Cldn-7 (green) in the colonic mucosa from the two groups of mice during injury and recovery periods; scale bars: 20 μm. The right panels show the quantification of the Lgr5 signal in each field of tissue at different time periods ( n = 3–4 per genotype). H Quantitative RT-PCR analysis of the indicated stem cell markers in colon tissues from Cldn-7 fl/fl ;Lgr5-CreERT2 and Cldn-7 fl/fl ;Lgr5-CreW mice at D5 and D12 ( n = 4–5 per genotype). Statistical data are presented as mean ± SEM. All p values were calculated using Student’s t -test (two-tailed) or Mann–Whitney nonparametric test; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns, not significant.

Article Snippet: A terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling (TUNEL) assay was performed using a one-step TUNEL apoptosis assay kit (Beyotime), according to the manufacturer’s instructions.

Techniques: Fluorescence, Staining, Expressing, RNAscope, Immunofluorescence, Quantitative RT-PCR, Flow Cytometry, TUNEL Assay, Disruption, In Situ, Hybridization, Two Tailed Test, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: Claudin-7 is essential for the maintenance of colonic stem cell homoeostasis via the modulation of Wnt/Notch signalling

doi: 10.1038/s41419-024-06658-x

Figure Lengend Snippet: A Representative bright field images of colonic organoids generated from Cldn-7 fl/fl ;Lgr5-CreERT2 mice treated with VEH or TAM at primary (P0) and first passage (P1); scale bars: 100 μm. B Statistical plots of the organoid-forming efficiency of primary cultures, average organoid sizes at day 8 of primary culture, and the replating efficiency at day 5 after first passage. All experiments were repeated three times with at least three mice in each group. C Representative image from Lgr5-EGFP + ISCs in VEH- and TAM-treated Cldn-7 fl/fl ;Lgr5-CreERT2 mice, visualised by fluorescent confocal microscopy 5 days after passage. Lgr5-EGFP immunofluorescence is shown in green (white arrowheads); scale bars: 100 μm. The right panel shows the quantification of Lgr5-EGFP + ISCs per organoid. D , E Representative images of colonic organoids in each group; samples were stained with HE, alongside Ki-67 (green) to detect proliferation, TUNEL (red) to detect apoptosis, Villin (red) to detect enterocytes, Muc2 (red) and Alcian blue to detect goblet cells, ChgA (red) to detect enteroendocrine cells, and Dclk1 (red) to detect tuft cells. Blue represents DAPI staining of nuclei; scale bars: 50 μm. Images are representative of at least three mice. F – H Statistical quantification of Ki-67, TUNEL, Villin, Muc2, ChgA, Dclk1-positive cells per organoid. Statistical data are presented as mean ± SEM. All p values were calculated using Student’s t -test (two-tailed) or Mann–Whitney nonparametric test; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns, not significant.

Article Snippet: A terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling (TUNEL) assay was performed using a one-step TUNEL apoptosis assay kit (Beyotime), according to the manufacturer’s instructions.

Techniques: Generated, Confocal Microscopy, Immunofluorescence, Staining, TUNEL Assay, Two Tailed Test, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: Claudin-7 is essential for the maintenance of colonic stem cell homoeostasis via the modulation of Wnt/Notch signalling

doi: 10.1038/s41419-024-06658-x

Figure Lengend Snippet: A Schematic diagram showing the injury and regeneration phases of experimental colitis in adult Cldn-7 fl/fl ;Lgr5-CreERT2 and Cldn-7 fl/fl ;Lgr5-CreW mice after TAM induction for 5 days. B Changes in body size of mice in each group after 3% DSS exposure. C Weight loss curves of Cldn-7 fl/fl ;Lgr5-CreERT2 and Cldn-7 fl/fl ;Lgr5-CreW mice during the 5 days of TAM induction and 7 days of DSS treatment ( n = 5 per genotype). D Representative images of colon length changes in mice following DSS induction (Injury) and 2 weeks of recovery (Regeneration). The right panel indicates the corresponding statistical data for colon length analysis ( n = 5 per genotype). E Representative histological HE (top), Ki-67 (middle), and TUNEL (bottom) staining images of colonic tissues from Cldn-7 fl/fl ;Lgr5-CreERT2 and Cldn-7 fl/fl ;Lgr5-CreW mice during acute inflammatory injury (7 days of DSS treatment) and regeneration (7 days of DSS followed by 14 days of regular water consumption). Arrowheads denote extensive mucosal defects and crypt destruction; scale bars: 50 μm. The right panels show the quantification of Ki-67-positive and TUNEL-positive cells in injured and regenerating epithelial tissue (three slides per sample; n = 3–4 per genotype). F , G Quantitative RT-PCR analysis of ISC-related marker genes in intact colonic tissues during intestinal inflammatory injury and regeneration, respectively, in each group of mice. All images are representative of at least three independent experiments. Statistical data are presented as mean ± SEM. All p values were calculated using Student’s t -test (two-tailed) or Mann–Whitney nonparametric test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns, not significant.

Article Snippet: A terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling (TUNEL) assay was performed using a one-step TUNEL apoptosis assay kit (Beyotime), according to the manufacturer’s instructions.

Techniques: TUNEL Assay, Staining, Quantitative RT-PCR, Marker, Two Tailed Test, MANN-WHITNEY

Journal: Cell death & disease

Article Title: The C-type lectin COLEC10 is predominantly produced by hepatic stellate cells and involved in the pathogenesis of liver fibrosis.

doi: 10.1038/s41419-023-06324-8

Figure Lengend Snippet: Fig. 2 The C-type lectins including Colec10 and Colec11 are highly expressed in quiescent hepatic stellate cells. A Expression profile of Acta2, Col1a1, Lrat, Colec10, Colec11, and Alb in the whole cell population. B Dotplot of the expression of Acta2, Col1a1, Lrat, Colec10, Colec11, and Alb in each cluster. C Trajectories of hepatic stellate cell activation. D Pseudotime inference of hepatic stellate cell activation. E Expression trend of genes along with increased pseudotime. F Gene expression profile of isolated hepatic stellate cells from healthy mice livers (Ctrl) and CCl4 challenged mice livers (24 h, 72 h, and 2w). The data are collected from GEO dataset GSE176042.

Article Snippet: Deparaffinized human liver sections were co-immunostained for COLEC10 (CUSABIO, China) and COL1A1 (Beyotime, China) with a two-step immunohistochemistry kit (Beyotime, China) and co-stained for ALB (Beyotime, China) and COLEC10 (CUSABIO, China) with a two-step immunofluorescence kit (Beyotime, China).

Techniques: Expressing, Activation Assay, Gene Expression, Isolation

Journal: Cell death & disease

Article Title: The C-type lectin COLEC10 is predominantly produced by hepatic stellate cells and involved in the pathogenesis of liver fibrosis.

doi: 10.1038/s41419-023-06324-8

Figure Lengend Snippet: Fig. 3 The C-type lectins including Colec11 and Colec10 are exclusively expressed in hepatic stellate cells and decreased in the fibrotic mice livers. A The landscape of cell clusters of isolated liver resident cells from CCL4-challenged mice livers. The data are collected from GEO dataset GSE212039. B The expression profile of cell marker genes in each cluster. C The HE staining and Masson trichrome staining of mice livers from mice treated with oil or CCl4 for 2 weeks or 4 weeks. Scale bar = 200 μm. D The RT-qPCR results of mRNA expression of mice livers treated with oil or CCl4 for 2 weeks or 4 weeks. E The immunofluorescence staining of HNF4a and COLEC10 of mice livers treated with oil or CCl4 for 2 weeks. Scale bar = 20 μm.

Article Snippet: Deparaffinized human liver sections were co-immunostained for COLEC10 (CUSABIO, China) and COL1A1 (Beyotime, China) with a two-step immunohistochemistry kit (Beyotime, China) and co-stained for ALB (Beyotime, China) and COLEC10 (CUSABIO, China) with a two-step immunofluorescence kit (Beyotime, China).

Techniques: Isolation, Expressing, Marker, Staining, Quantitative RT-PCR

Journal: Cell death & disease

Article Title: The C-type lectin COLEC10 is predominantly produced by hepatic stellate cells and involved in the pathogenesis of liver fibrosis.

doi: 10.1038/s41419-023-06324-8

Figure Lengend Snippet: Fig. 5 The expression of COLEC10 in human fibrotic livers. A The cirrhotic human liver section was co-stained for ALB and COLEC10. Magnification 400 ´ . B The human liver sections collected from patients with hepatitis or cirrhosis were stained for COLEC10 or COL1A1. Scale bar = 100 μm. C The positive area percentage of IHC images from patients with hepatitis or cirrhosis were quantified and compared. D The expression of interested genes in bulk RNA sequencing data of human liver samples from a cohort of patients diagnosed as liver fibrosis. The data are collected from GEO dataset GSE193066.

Article Snippet: Deparaffinized human liver sections were co-immunostained for COLEC10 (CUSABIO, China) and COL1A1 (Beyotime, China) with a two-step immunohistochemistry kit (Beyotime, China) and co-stained for ALB (Beyotime, China) and COLEC10 (CUSABIO, China) with a two-step immunofluorescence kit (Beyotime, China).

Techniques: Expressing, Staining, RNA Sequencing

Journal: Cell death & disease

Article Title: The C-type lectin COLEC10 is predominantly produced by hepatic stellate cells and involved in the pathogenesis of liver fibrosis.

doi: 10.1038/s41419-023-06324-8

Figure Lengend Snippet: Fig. 6 The effect of COLEC10 overexpression in LX-2 cells. A Protein expression of LX-2 cells transfected with control vector or COLEC10 over-expression vector. B The volcano plot of differentially expressed genes between vector transfected LX-2 cells and COLEC10 overexpression LX-2 cells in the RNA sequencing data. C The heatmap of scaled expression of differentially expressed genes in each sample of vector transfected LX-2 cells and COLEC10 overexpression LX-2 cells. D Gene Ontology (GO) classification of differentially expressed genes between samples. E The Kyoto Encyclopedia of Genes and Genomes (KEGG) classification of differentially expressed genes between samples. F The Reactome annotation pathway of differentially expressed genes.

Article Snippet: Deparaffinized human liver sections were co-immunostained for COLEC10 (CUSABIO, China) and COL1A1 (Beyotime, China) with a two-step immunohistochemistry kit (Beyotime, China) and co-stained for ALB (Beyotime, China) and COLEC10 (CUSABIO, China) with a two-step immunofluorescence kit (Beyotime, China).

Techniques: Over Expression, Expressing, Transfection, Control, Plasmid Preparation, RNA Sequencing

Journal: Cell death & disease

Article Title: The C-type lectin COLEC10 is predominantly produced by hepatic stellate cells and involved in the pathogenesis of liver fibrosis.

doi: 10.1038/s41419-023-06324-8

Figure Lengend Snippet: Fig. 7 The immune regulation and extracellular matrix alteration associated genes are increased in the COLEC10 over-expression LX-2 cells. A The Reactome pathway analysis of upregulated genes in the COLEC10 over-expression LX-2 cells. B The Reactome pathway analysis of downregulated genes in the COLEC10 over-expression LX-2 cells. C The heatmap of scaled expression of genes in the interleukin-4 and interleukin-13 signaling pathway and the cell surface interactions at the vascular wall pathway between the COLEC10 over-expression LX-2 cells and the vector control LX-2 cells. D The mRNA expression of COL1A1, COL1A2, COL3A1, MMP2 and ACTA2 in the COLEC10 over-expression LX-2 cells and the vector control LX-2 cells. The GAPDH and β-Actin were used as loading control. E The protein expression of COL1A1 and ACTA2 in the COLEC10 over- expression LX-2 cells and the vector control LX-2 cells. F The quantificiation of protein expression of COL1A1 and ACTA2 in the COLEC10 over- expression LX-2 cells and the vector control LX-2 cells.

Article Snippet: Deparaffinized human liver sections were co-immunostained for COLEC10 (CUSABIO, China) and COL1A1 (Beyotime, China) with a two-step immunohistochemistry kit (Beyotime, China) and co-stained for ALB (Beyotime, China) and COLEC10 (CUSABIO, China) with a two-step immunofluorescence kit (Beyotime, China).

Techniques: Over Expression, Expressing, Plasmid Preparation, Control

Journal: Cell death & disease

Article Title: The C-type lectin COLEC10 is predominantly produced by hepatic stellate cells and involved in the pathogenesis of liver fibrosis.

doi: 10.1038/s41419-023-06324-8

Figure Lengend Snippet: Fig. 8 The serum COLEC10 is increased in patients with chronic liver disease. A The comparison of serum COLEC10 concentration between healthy donors and patients with chronic liver diseases (CLD). B The serum concentration of COLE10 across different Child-Pugh stages of CLD patients. C The correlation plot of COLEC10 with other serum markers and clinical features of patients with CLD. D The linear regression of COLEC10 to D-dimer in the patients with CLD.

Article Snippet: Deparaffinized human liver sections were co-immunostained for COLEC10 (CUSABIO, China) and COL1A1 (Beyotime, China) with a two-step immunohistochemistry kit (Beyotime, China) and co-stained for ALB (Beyotime, China) and COLEC10 (CUSABIO, China) with a two-step immunofluorescence kit (Beyotime, China).

Techniques: Comparison, Concentration Assay

Journal: Cell death & disease

Article Title: The C-type lectin COLEC10 is predominantly produced by hepatic stellate cells and involved in the pathogenesis of liver fibrosis.

doi: 10.1038/s41419-023-06324-8

Figure Lengend Snippet: Fig. 9 AAV transduced COLEC10 overexpression contributes to ECM alteration and angiogenesis in CCl4 challenged mice livers. A Illustration of AAV transduction and CCl4 administration of mice models. B mRNA expression of genes of mice livers in AAV control group (Ctrl) and AAV overexpression group (OE).

Article Snippet: Deparaffinized human liver sections were co-immunostained for COLEC10 (CUSABIO, China) and COL1A1 (Beyotime, China) with a two-step immunohistochemistry kit (Beyotime, China) and co-stained for ALB (Beyotime, China) and COLEC10 (CUSABIO, China) with a two-step immunofluorescence kit (Beyotime, China).

Techniques: Over Expression, Transduction, Expressing, Control

Journal: Cellular and Molecular Life Sciences

Article Title: Vascular adhesion protein-1 defines a unique subpopulation of human hematopoietic stem cells and regulates their proliferation

doi: 10.1007/s00018-021-03977-6

Figure Lengend Snippet: VAP-1 expression on human BM and CB HSC. a Flow cytometric identification of HSC in human BM. The plots show the sequential gating strategy for identifying Lineage − CD34 + CD38 − CD90 + CD45RA − CD49f + HSC (gate P-5). b Staining of gate P-5 HSC using the anti-VAP-1 antibody JG-2 or an isotype-matched negative control antibody. c Identification of human CB HSC from FACS sorted CD34 + CB cells. d CB HSC (gate P-5) analyzed for VAP-1 expression as in b . e Expression of VAP-1 on FACS-sorted CD34 + CB cells using immunofluorescence staining of cytospin slides. The arrow points to a VAP-1 + cell. The inset shows a negative control staining. Scale bar 50 µm. In b and d , the numbers indicate the percentages of VAP-1 + cells (BM 19.5 ± 2%, n = 5; CB 51.8 ± 8.7%, n = 10

Article Snippet: CD34 + cells from human umbilical CB were isolated via a two-step procedure using Ficoll-Plaque gradient centrifugation (Amersham Pharmacia Biotech, Uppsala, Sweden) and EasySep Human Cord Blood CD34 Positive Selection Kit II (STEMCELL Technologies).

Techniques: Expressing, Staining, Negative Control, Immunofluorescence

Journal: Cellular and Molecular Life Sciences

Article Title: Vascular adhesion protein-1 defines a unique subpopulation of human hematopoietic stem cells and regulates their proliferation

doi: 10.1007/s00018-021-03977-6

Figure Lengend Snippet: VAP-1 − and VAP-1 + HSC from adult human BM are transcriptionally different. a Population-distance analysis of VAP-1 − and VAP-1 + HSC (Lin − CD34 + CD38 − CD45RA − CD90 + CD49f + cells) presented as a multidimensional scaling plot of the bulk RNAseq data in two dimensions. Each point represents a single biological replicate. b A heatmap of genes differentially expressed in VAP-1 − and VAP-1 + HSC. Colors represent the relative expression of a given gene in comparison with the median of all samples. The heat map color scheme with the corresponding log2-values is given, n = 4 in both groups. c A volcano plot showing the distribution of gene expression changes in VAP-1 − and VAP-1 + HSC. Genes were plotted as − log10( p -value) on the y axis against the log2(fold-change) on the x axis. d Quantitative real-time polymerase chain reaction (qPCR) of selected differentially expressed genes in VAP-1 + and VAP-1 − HSC. The expression intensity was normalized with the expression level of β2 microglobulin ( B2M). Two samples independent from those used in RNAseq were analyzed ( p -value < 0.05). e Gene set enrichment analysis (GSEA) of the HSC signature genes in VAP-1 − and VAP-1 + HSC. Normalized enrichment score (NES). f Gene ontology (GO) molecular signaling pathway analysis for DEGs. Shown are the top five pathways (FDR < 0.02 and > 50 observed genes. g GSEA for ROS production, hallmark, metabolism and detoxification. NES and FDR-q values are shown. h GSEA for p53, MAPK, p38, FOXO, HIF, AKT and ATM signaling pathways. NES and FDR-q values are shown. i The enrichment plot of the p53 signaling pathway. j A protein–protein interaction (gray lines, the thickness of which represents confidence and strength of the association) network of the enriched genes of the p53 signaling pathway in VAP-1 + HSC. Proteins are shown as spheres

Article Snippet: CD34 + cells from human umbilical CB were isolated via a two-step procedure using Ficoll-Plaque gradient centrifugation (Amersham Pharmacia Biotech, Uppsala, Sweden) and EasySep Human Cord Blood CD34 Positive Selection Kit II (STEMCELL Technologies).

Techniques: Expressing, Comparison, Gene Expression, Real-time Polymerase Chain Reaction, Protein-Protein interactions

Journal: Cellular and Molecular Life Sciences

Article Title: Vascular adhesion protein-1 defines a unique subpopulation of human hematopoietic stem cells and regulates their proliferation

doi: 10.1007/s00018-021-03977-6

Figure Lengend Snippet: Inhibition of VAP-1 expands BM and CB HSC in in vitro cultures. a The number of colonies arising from single-cell sorted VAP-1 + and VAP-1 − CB. b The number of colonies from ( a ) re-plated and cultured for another 12–14 days. c The number of colonies from ( b ) counted by microscopy ( p -value < 0.01). d Characteristics and number of CFU-E, CFU-GM or CFU-MIX colonies. a – d ; CB collected from 60 donors. e Number of BM HSC in CFU assays in presence or absence of VAP-1 inhibition. Five hundred human BM-derived CD34 + cells were cultured under CFU conditions in presence of LJP-1586 (0.5 µM) or vehicle. n = two donors, triplicate cultures ( p -value < 0.05). f Kinetic analyses of sorted CD38 − CD34 + CB-derived cells cultured with different concentrations of LJP-1586 for 21 days. Data from one representative donor out of 3 are shown. g Effect of LJP-1586 (1 µM) on FACS-sorted CD38 − CD34 + CB cells cultured in vitro for 15 d, n = 4 donors ( p -value < 0.05). h Analyses of CD34 + CD38-CD45RA − CD90 + CD49f + HSC expansion subsequent to 15d LJP-1586 treatment measured as fold expansion and CFUs ( n = 4) ( p -value < 0.05)

Article Snippet: CD34 + cells from human umbilical CB were isolated via a two-step procedure using Ficoll-Plaque gradient centrifugation (Amersham Pharmacia Biotech, Uppsala, Sweden) and EasySep Human Cord Blood CD34 Positive Selection Kit II (STEMCELL Technologies).

Techniques: Inhibition, In Vitro, Cell Culture, Microscopy, Derivative Assay

Journal: Cellular and Molecular Life Sciences

Article Title: Vascular adhesion protein-1 defines a unique subpopulation of human hematopoietic stem cells and regulates their proliferation

doi: 10.1007/s00018-021-03977-6

Figure Lengend Snippet: Inhibition of VAP-1 increases the engraftment potential of CB and BM HSC in NBSGW mice. a The experimental set up for in vivo analysis of mononuclear CB cells with VAP-1 inhibitor. b Gating strategy of harvested BM cells from recipients at the end of the experiment by flow cytometry using antibodies against mouse CD45, human CD45, human CD34 and human CD38. c Percentages of cells isolated from BM of transplanted mice ( n = 7) within the different gates defined in ( b ) ( p -value < 0.05). d The experimental scheme for in vivo analyses of VAP-1 – and VAP-1 +/lo HSC in the presence of VAP-1 inhibitor. e Batch sorting and the frequencies of VAP-1 − and VAP-1 +/lo HSC from human BM in the CD34 + gate. f In vivo engraftment of VAP-1 − and VAP-1 +/lo human BM cells in non-irradiated NBSGW mice after 42 days. Representative flow cytometric plots (percentages of engrafted cells) and quantification from each group ( n = 3 for each group). The cut-off value for engraftment was set as 0.1

Article Snippet: CD34 + cells from human umbilical CB were isolated via a two-step procedure using Ficoll-Plaque gradient centrifugation (Amersham Pharmacia Biotech, Uppsala, Sweden) and EasySep Human Cord Blood CD34 Positive Selection Kit II (STEMCELL Technologies).

Techniques: Inhibition, In Vivo, Flow Cytometry, Isolation, Irradiation

Journal: Cellular and Molecular Life Sciences

Article Title: Vascular adhesion protein-1 defines a unique subpopulation of human hematopoietic stem cells and regulates their proliferation

doi: 10.1007/s00018-021-03977-6

Figure Lengend Snippet: LJP-1586 treatment increases the number of HSC in BM of mice. a Percentages of HSC within gate 6 (shown in Fig. a) in the groups treated by inhibitor LJP-1586 and control PBS ( p -value < 0.05). b FACS profiles showing gradual enrichment of HSC using a different set of HSC markers as used in Fig. a (Lin-CD41-Sca-1 + c-Kit + CD48-CD150 +) and the bars show the percentage of HSC from gate P-6 ( p -value < 0.01). c CFU assay using peripheral blood (PB) or BM cells isolated after the 7-day treatment with LJP-1586 ( n = 4). d CFU assay using PB or BM cells isolated after the 42-day treatment with LJP-1586 ( n = 10) ( p -value < 0.05). e Analysis of BM derived subtypes of CFUs shown in d . BFU-E Burst forming erythroid unit, CFUM macrophage colony forming unit, CFU-G granulocyte colony forming unit, CFUGM granulocyte/macrophage colony forming unit, CFU-GEMM granulocyte/erythroid/macrophage/megakaryocyte colony forming unit. Student t -test was applied ( p -value < 0.01)

Article Snippet: CD34 + cells from human umbilical CB were isolated via a two-step procedure using Ficoll-Plaque gradient centrifugation (Amersham Pharmacia Biotech, Uppsala, Sweden) and EasySep Human Cord Blood CD34 Positive Selection Kit II (STEMCELL Technologies).

Techniques: Control, Colony-forming Unit Assay, Isolation, Derivative Assay