Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Detection of β-catenin, E- and N-cadherins and the cellular localization of the proteins in chronic hepatitis C (CH-C), hepatocellular carcinoma (HCC) and normal liver (control)

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Control

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Immunohistochemical ( A – C ) and hybridocytochemical localization of β-catenin ( D ) in liver. Membranous/ cytoplasmic localization of β-catenin in liver with chronic hepatitis C ( A ), membranous localization of β-catenin in hepatocellular carcinoma ( B ) and in control liver ( C ); mRNA for β-catenin in cytoplasm and cell nuclei in HCC fragment ( D ). Immunocytochemical detection of E-cadherin ( E – F ) and N-cadherin ( H ) in liver. Predominantly membranous localization of E-cadherin in liver with chronic hepatitis C ( E ), hepatocellular carcinoma ( F ) and control liver ( G ). Membrano-cytoplasmic localization of N-cadherin in fragment of hepatocellular carcinoma ( H ). Immunohistochemistry ( A – C , E – H ) technique and hybridization in situ method ( D ). Hematoxylin counterstained. Bar = 40 μm

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Immunohistochemical staining, Control, Immunohistochemistry, Hybridization, In Situ

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Comparison of quantitatively assessed β-catenin, E-cadherin and N-cadherin immunoexpression (% of the IHC reaction area in analysed area of liver parenchyma) in chronic hepatitis C (CH-C), hepatocellular carcinoma (HCC) and normal liver (C)

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Comparison

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Comparative immunoexpression of β-catenin, E-cadherin and N-cadherin in liver with chronic hepatitis C (CH-C), hepatocellular carcinoma (HCC) and normal organ (control) ***p (level of significance) value < 0.001, *p < 0.05.

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Control

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Tissue expression of β-catenin, E-cadherin and N-cadherin (mean % of IHC reaction area ± SD) as related to grading and staging in chronic hepatitis C (CH-C) group

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Expressing

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Values of Spearman’s rank coefficient between expression of β-catenin, E-cadherin, N-cadherin (% IHC reaction area per area of hepatic parenchyma) and clinicopathological data in CH-C group

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Expressing, Infection

Journal: Immunity

Article Title: Notch4 signaling limits regulatory T-cell-mediated tissue repair and promotes severe lung inflammation in viral infections

doi: 10.1016/j.immuni.2021.04.002

Figure Lengend Snippet:

Article Snippet: Antibodies against the following human antigens were used: CD3 (HIT3a, catalog no: 300318, 1:200,Biolegend), CD4 (RPA-T4, catalog no: 300530, 1:200,Biolegend), Foxp3 (PCH-101,catalog no: 48-4776-42,56-4716-41, 1:100 Thermofisher), Helios (22F6, catalog no: 47-9883-42 1:100, Thermofisher), Notch1 (HMN1-519, catalog no: 566023, 1:300, BD PharMingen), Notch2 (HMN2-25, catalog no: 742291, 1:300, BD PharMingen), Notch3 (HMN3-21, catalog no: 744828, 1:300, BD PharMingen), Notch4 (HMN4-2, Catalogue no: 563269, 1:300, BD PharMingen), Yap1 (D8H1X, Catalogue no: 14729S, 1:100, Cell Signaling Technology), beta-Catenin (196624, Catalogue no: IC13292V, 1:200, R&D system), CD25 (BC96, Catalogue no: 51-025-941 1:300, Thermofisher), IL-4 (MP4-25D2, Catalogue no: 500828 1:200, Biolegend), IL-13 (JES10-5A2, Catalogue no: 501912, 1:200, Biolegend), CCR4 (L291H4, Catalogue no: 359417 1:300, Biolegend), CXCR3 (G025H7, Catalogue no: 353708, 1:300 Biolegend), CRTH2 (BM16, Catalogue no: 350108 1:300, Biolegend), CD127 (A019D5, Catalogue no: 351320 1:300, Biolegend), IFNγ (4S.B3, Catalogue no: 560741 1:200, BD Biosciences) and Amphiregulin polyclonal antibody (orb7378 1:200, Biorbyt), CD196 (G034E3, Catalogue no: 353423, 1:300 Biolegend).

Techniques: Recombinant, Adjuvant, Enzyme-linked Immunosorbent Assay, Control, Blocking Assay, Mutagenesis, Software

Journal: American Journal of Physiology - Renal Physiology

Article Title: Vimentin expression is required for the development of EMT-related renal fibrosis following unilateral ureteral obstruction in mice

doi: 10.1152/ajprenal.00340.2017

Figure Lengend Snippet: Vimentin (vim) knockout mice undergoing unilateral ureteral obstruction (UUO) demonstrate altered β-catenin, E-cadherin, and keratin staining on Western blotting of whole kidneys 1, 2, and 4 wk following UUO. Whole kidneys from wild-type (WT) and vim −/− were recovered and homogenized in 10 mM Tris (pH 7.4) buffer without SDS. This SDS-deficient lysis buffer was used to minimize solubilization of membrane-associated proteins and to detect total cytosolic β-catenin levels (48). A: immunoblotting with anti-E-cadherin, -vim, -β-catenin, and -pan-keratin antibodies was performed. Vim was detected 1 wk following UUO in control mice. Nonmembrane-bound β-catenin was detected 2 wk following UUO in control mice but 1 wk after UUO in vim −/− mice. E-cadherin signaling was diminished when comparing vim −/− mice vs. control mice, whereas keratin staining increased from 1 to 2 wk in vim −/− mice and decreased in control mice. Actin staining was used as loading controls. Molecular weight is in kDa. B: Western blot staining was normalized against unligated control kidneys. Vim staining increased ~fourfold over control kidneys and peaks at 2 wk (~eightfold increase; P > 0.05). Nonmembrane-bound β-catenin increases fourfold over control kidneys 1 wk following UUO and peaks at 4 wk in vim −/− mice although β-catenin levels also increase in control kidneys over time following UUO. E-cadherin levels decrease in both WT and vim −/− kidneys 1 and 2 wk following UUO. Keratin staining increases in both vim −/− and WT mice, but keratin levels are higher in vim −/− mice. n = 4. Error bars = standard error, *P < 0.05.

Article Snippet: After blocking, the samples were exposed to the following primary antibodies: goat polyclonal beta-catenin (1:50, R&D Systems, Minneapolis, MN), rabbit polyclonal vim (1:50, Santa Cruz Biotechnology, Dallas, TX), and goat polyclonal E-cadherin (1:50, R&D Systems), all diluted in blocking solution.

Techniques: Knock-Out, Staining, Western Blot, Lysis, Membrane, Molecular Weight

Journal: American Journal of Physiology - Renal Physiology

Article Title: Vimentin expression is required for the development of EMT-related renal fibrosis following unilateral ureteral obstruction in mice

doi: 10.1152/ajprenal.00340.2017

Figure Lengend Snippet: β-catenin mRNA production is not affected in vimentin (vim) −/− mice undergoing unilateral ureteral obstruction (UUO). qPCR primers for vim, β-catenin, and keratin 8 were designed and utilized to probe wild-type (WT) and vim −/− kidneys 1, 2, and 4 wk following UUO. qPCR readouts were normalized to ribosome 18 s (Rn18s) to account for variability in loading and then expressed as fold changes of the internal control, nonligated right kidney, for each individual mouse that was analyzed. Relative mRNA expression to Rn18s shows increase in vim expression at 1 and 2 wk following UUO in WT kidneys, whereas vim expression is detected relative to Rn18s 4 wk following UUO in the unligated WT control kidney (A). β-catenin RNA expression is not statistically significant among all kidneys 1, 2, and 4 wk following UUO (B). Keratin 8 mRNA levels are also increased 1 wk post-UUO in WT ligated kidneys. At 4 wk, there is an increase of keratin 8 when compared with the other samples (C). mRNA levels were then normalized against nonligated right kidney (taken as 1). Histogram shows the mRNA levels of ligated left kidney. Vim expression was detected 1 and 2 wk following UUO and decreased sharply 4 wk post-UUO (D). There is no statistical significance in the detection of β-catenin mRNA 1, 2, and 4 wk post-UUO between WT and vim −/− mice (E). Keratin 8 mRNA levels are slightly higher in vim −/− mice 2 and 4 wk post-UUO (F). Data are representative of n = 4 independent experiments. Error bars = Standard error; *P < 0.05.

Article Snippet: After blocking, the samples were exposed to the following primary antibodies: goat polyclonal beta-catenin (1:50, R&D Systems, Minneapolis, MN), rabbit polyclonal vim (1:50, Santa Cruz Biotechnology, Dallas, TX), and goat polyclonal E-cadherin (1:50, R&D Systems), all diluted in blocking solution.

Techniques: Expressing, RNA Expression

Journal: American Journal of Physiology - Renal Physiology

Article Title: Vimentin expression is required for the development of EMT-related renal fibrosis following unilateral ureteral obstruction in mice

doi: 10.1152/ajprenal.00340.2017

Figure Lengend Snippet: Vimentin (vim) −/− mice undergoing unilateral ureteral obstruction (UUO) show altered cellular localization of β-catenin and E-cadherin in proximal renal tubules. OCT-embedded kidneys following UUO underwent immunofluorescence with antibodies directed against vim, β-catenin. DAPI was used for nuclear staining. All images were obtained on with a Leica DMI4000 B confocal microscope and analyzed using Leica Advanced Fluorescence Application Suite. Wild-type (WT) kidneys 1 wk post-UUO have an increased expression of vim in the cytoplasm of proximal renal tubular cells (C). However, β-catenin staining colocalizes with DNA in the nucleus (D, arrow). Vim −/− kidneys 1-wk post-UUO kidneys result in β-catenin signals more prominently in the cytoplasm (F, H; arrow), with no detectable vim staining (G). Mander’s colocalization coefficients calculation reveals a statically significant increase in colocalization between DAPI and β-catenin staining in WT vs. vim −/− mice (0.34 vs. 1.62). Size bars = 10 μm. *P < 0.05.

Article Snippet: After blocking, the samples were exposed to the following primary antibodies: goat polyclonal beta-catenin (1:50, R&D Systems, Minneapolis, MN), rabbit polyclonal vim (1:50, Santa Cruz Biotechnology, Dallas, TX), and goat polyclonal E-cadherin (1:50, R&D Systems), all diluted in blocking solution.

Techniques: Immunofluorescence, Staining, Microscopy, Fluorescence, Expressing

Journal: American Journal of Physiology - Renal Physiology

Article Title: Vimentin expression is required for the development of EMT-related renal fibrosis following unilateral ureteral obstruction in mice

doi: 10.1152/ajprenal.00340.2017

Figure Lengend Snippet: Vimentin (vim) mediates transforming growth factor (TGF)-β-induced epithelial to mesenchymal transition (EMT) of human renal tubular epithelial cells (A). Sequence of shRNA to human vim. shRNA specially targeting (shVIM) downregulated vim expression efficiently in human proximal renal tubular (HK-2) cells (B). Two and 3 days following induction with TGF-β, control HK-2 cell lines express vim, whereas shVIM cell lines (VIM) do not express vim in appreciable amounts. All lanes were normalized to β-actin to ensure proper loading. Time lapse phase contrast microscopy was used to visualize wound healing in control cells (shCON) and shVIM cells following TGF-β induction (C). Quantification of cellular migration was carried out by measuring the changes in the wound area after 24 h (D). There was a ~30% reduction in migration following EMT induction in shVIM cell lines. Experiments were performed in triplicate and P values calculated. Immunoblotting using antibodies against vim, pan-keratin, and EMT markers, including zona occludens (ZO)-1, N-cadherin, E-cadherin, and β-catenin was carried out on shVIM HK-2 cells following TGF-β induction (E). Signals were normalized to GAPDH to account for variability in loading and then expressed as fold changes of the negative control, shCON. shVIM HK-2 cells demonstrate “sheet-like” motility following TGF-β stimulation. Vim-silenced HK-2 cells were treated with TGF-β and then underwent wound healing assay. Time lapse phase contrast microscopy for 24 h demonstrate sheet-like movement when compared with control cell lines. Time lapse assembled from 1-h images over 24-h period. Supplemental Material for this article is available online at the Journal website.

Article Snippet: After blocking, the samples were exposed to the following primary antibodies: goat polyclonal beta-catenin (1:50, R&D Systems, Minneapolis, MN), rabbit polyclonal vim (1:50, Santa Cruz Biotechnology, Dallas, TX), and goat polyclonal E-cadherin (1:50, R&D Systems), all diluted in blocking solution.

Techniques: Sequencing, shRNA, Expressing, Microscopy, Migration, Western Blot, Negative Control, Wound Healing Assay

Journal: Nature Communications

Article Title: Clonal evolution patterns in acute myeloid leukemia with NPM1 mutation

doi: 10.1038/s41467-019-09745-2

Figure Lengend Snippet: Flow cytometry of total and active CTNNB1 in diagnosis samples of NPM1 mut loss and NPM1 mut persistent pts. Higher expression of a CTNNB1 total, b CTNNB1 8e7 in 3 NPM1 mut persistent diagnosis samples compared to 4 NPM1 mut loss diagnosis samples, and c mean fluorescent intensity (MFI) including all data points, data is presented as mean ± s.d

Article Snippet: We used a primary antibody against total CTNNB1 (anti-h-beta-Catenin, APC-conjugated, #IC13292A, R&D Systems) or active CTNNB1 (anti-active-b-Catenin, clone 8E7, #05-665-25UG, Merck Millipore) followed by a secondary Alexa488 labeled antibody (donkey anti-mouse, Invitrogen # R37114).

Techniques: Flow Cytometry, Biomarker Discovery, Expressing

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: β-Catenin , REA480 , 50 , 130-123-546 , FITC (APC, PE) , Miltenyi Biotec.

Techniques: Imaging

Journal: PLoS Pathogens

Article Title: Memory like NK cells display stem cell like properties after Zika virus infection

doi: 10.1371/journal.ppat.1009132

Figure Lengend Snippet: ( A ) Hallmark pathway gene sets analysis in FACS purified CD27 + memory like and non-memory CD27 - NK cells a month post ZIKV infection. RNA-seq data are from 3 biological replicates for each group. ( B ) ATAC-seq tracks for selected loci of Wnt/β-catenin pathway genes in CD27 + memory like (green) and non-memory CD27 - NK cells (blue). ( C-D ) GSEA enrichment plots ( C ) and heat maps ( D ) for Wnt/β-catenin pathway ( C left ) and canonical Wnt targets ( C right ) among CD27 + memory like and non-memory CD27 - NK gene sets. (E - G ) Immunofluorescence images of FACS purified CD27 + memory like and non-memory CD27 - NK cells, isolated a month post ZIKV infection, and stained for TCF-1 (yellow) and β-catenin (red) show co-localization of both molecules in CD27 + cells. Scale bar corresponds to 10 μm. Top-CD27 + NK cells, Bottom-CD27 - NK cells. Data are representative of 2 independent experiments. ( G ) Relative fluorescent intensity of TCF-1 in CD27 + memory like and non-memory CD27 - NK. Data are representative of 2 independent experiments (n = 4 per experiment). ( H ) Transcription factor motif enrichment analysis of ATAC-seq from CD27 + memory like NK cells. ( I ) % of target sequences with TCF-1 motifs in genomic regions of CD27 + memory like and non-memory CD27 - NK cells. Chromatin opening by ATAC-seq ( J ) and gene expression by RNA-seq ( K ) of selected TCF-1 target genes in CD27 + memory like NK cells. Fold change values for peaks are plotted in ( J ). Mean ± s.d. two-sided Student’s t-test, *** P ≤ 0.001.

Article Snippet: This was followed by antibody staining step and cells were incubated with TCF-1-PE (4:100) (S33-966, BD Biosciences, #564217) and β-catenin-APC (4:100) (REA480, Miltenyi Biotech, #130124453) antibodies in PBS with 0.5% BSA for 45 min. Then, cells were washed with PBS containing 0.5% BSA, pelleted and subjected to cytospin for 2 minutes and taken on the slides.

Techniques: Purification, Infection, RNA Sequencing, Immunofluorescence, Isolation, Staining, Gene Expression