Journal: Cell Stem Cell

Article Title: Constitutively Active SMAD2/3 Are Broad-Scope Potentiators of Transcription-Factor-Mediated Cellular Reprogramming

doi: 10.1016/j.stem.2017.10.013

Figure Lengend Snippet:

Article Snippet: The following antibodies were used with indicated dilution: ICAM1-biotin conjugate (1:100, eBioscience, #13-0541), CD44-allophycocyanin (APC) conjugate (1:300, eBioscience, #17-0441), E-CADHERIN-eFluor 660 (1:300, eBioscience, #50-3249-82), MEFSK4-biotin conjugate (1:100, Miltenyi, 130-101-875), CD47-biotin conjugate (1:100, BioLegend, 127505), CD73-Alexa Fluor 647 (1:300, BD Biosciences, 561543) and CD104-APC (1:300, Miltenyi, 130-106-924) and Strepdavidin PE-Cy7 (1:1500, eBioscience, #25-4317).

Techniques: Recombinant, Software

Journal: BMC Cancer

Article Title: Identification and functional characterization of genes associated with lipopolysaccharide in lung adenocarcinoma

doi: 10.1186/s12885-025-15387-z

Figure Lengend Snippet: Expression of ITGB4 in Lung Adenocarcinoma Cells and Tissues. A Comparison of ITGB4 expression between 483 LUAD samples and 347 normal samples. B K–M survival curves for high and low ITGB4 expression groups. C – D ITGB4 expression levels in normal lung epithelial cell lines and LUAD cell lines analyzed by qRT-PCR and WB. E WB analysis of ITGB4 expression in 12 paired LUAD tissues. F IHC detection of ITGB4 expression in LUAD tissues. * p < 0.05, ** p < 0.01

Article Snippet: The membranes were blocked with 5% non-fat milk and incubated overnight at 4 °C with the following primary antibodies: ITGB4 (Proteintech, 21738-1-AP), GAPDH (Proteintech, 10494-1-AP), MMP2 (Proteintech, 10373-2-AP), MMP9 (Proteintech, 10375-2-AP), p53 (Proteintech, 10442-1-AP), Caspase-3 (Cell Signaling Technology, 9664 T), and Bcl-2 (Cell Signaling Technology, 4223 T).

Techniques: Expressing, Comparison, Quantitative RT-PCR

Journal: BMC Cancer

Article Title: Identification and functional characterization of genes associated with lipopolysaccharide in lung adenocarcinoma

doi: 10.1186/s12885-025-15387-z

Figure Lengend Snippet: Knockdown of ITGB4 Inhibits Proliferation, Migration, and Invasion of Lung Adenocarcinoma Cells A.Efficiency of ITGB4 knockdown in A549 and PC9 cells validated by qRT-PCR and WB. B.Cell proliferation assessed using the CCK-8 assay. C.Cell migration evaluated by wound healing assay. D.Expression levels of migration-related proteins (MMP2, MMP9) detected by WB. E. Transwell assay used to assess cell migration and invasion abilities. *** p < 0.001, **** p < 0.0001

Article Snippet: The membranes were blocked with 5% non-fat milk and incubated overnight at 4 °C with the following primary antibodies: ITGB4 (Proteintech, 21738-1-AP), GAPDH (Proteintech, 10494-1-AP), MMP2 (Proteintech, 10373-2-AP), MMP9 (Proteintech, 10375-2-AP), p53 (Proteintech, 10442-1-AP), Caspase-3 (Cell Signaling Technology, 9664 T), and Bcl-2 (Cell Signaling Technology, 4223 T).

Techniques: Knockdown, Migration, Quantitative RT-PCR, CCK-8 Assay, Wound Healing Assay, Expressing, Transwell Assay

Journal: BMC Cancer

Article Title: Identification and functional characterization of genes associated with lipopolysaccharide in lung adenocarcinoma

doi: 10.1186/s12885-025-15387-z

Figure Lengend Snippet: ITGB4 Knockdown Induces G1 Phase Arrest and Promotes Apoptosis. A Effects of ITGB4 knockdown on the cell cycle of PC9 cells. B Effects of ITGB4 knockdown on apoptosis in PC9 cells. C Changes in the expression of apoptosis-related proteins (Caspase-3, P53, Bcl-2) in PC9 cells. *** p < 0.001

Article Snippet: The membranes were blocked with 5% non-fat milk and incubated overnight at 4 °C with the following primary antibodies: ITGB4 (Proteintech, 21738-1-AP), GAPDH (Proteintech, 10494-1-AP), MMP2 (Proteintech, 10373-2-AP), MMP9 (Proteintech, 10375-2-AP), p53 (Proteintech, 10442-1-AP), Caspase-3 (Cell Signaling Technology, 9664 T), and Bcl-2 (Cell Signaling Technology, 4223 T).

Techniques: Knockdown, Expressing

Journal: BMC Cancer

Article Title: Identification and functional characterization of genes associated with lipopolysaccharide in lung adenocarcinoma

doi: 10.1186/s12885-025-15387-z

Figure Lengend Snippet: Functional and Pathway Analysis Regulated by ITGB4. A Volcano plot of differentially expressed genes. B GO enrichment analysis. C – D KEGG enrichment analysis

Article Snippet: The membranes were blocked with 5% non-fat milk and incubated overnight at 4 °C with the following primary antibodies: ITGB4 (Proteintech, 21738-1-AP), GAPDH (Proteintech, 10494-1-AP), MMP2 (Proteintech, 10373-2-AP), MMP9 (Proteintech, 10375-2-AP), p53 (Proteintech, 10442-1-AP), Caspase-3 (Cell Signaling Technology, 9664 T), and Bcl-2 (Cell Signaling Technology, 4223 T).

Techniques: Functional Assay

Journal: BMC Cancer

Article Title: Identification and functional characterization of genes associated with lipopolysaccharide in lung adenocarcinoma

doi: 10.1186/s12885-025-15387-z

Figure Lengend Snippet: ITGB4 Promotes Tumor Growth In Vivo. A Xenograft model in nude mice. B Tumor volume measurement. C Tumor weight evaluation. D ITGB4 expression levels detected in the orthotopic lung cancer model by IHC. ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The membranes were blocked with 5% non-fat milk and incubated overnight at 4 °C with the following primary antibodies: ITGB4 (Proteintech, 21738-1-AP), GAPDH (Proteintech, 10494-1-AP), MMP2 (Proteintech, 10373-2-AP), MMP9 (Proteintech, 10375-2-AP), p53 (Proteintech, 10442-1-AP), Caspase-3 (Cell Signaling Technology, 9664 T), and Bcl-2 (Cell Signaling Technology, 4223 T).

Techniques: In Vivo, Expressing, Paraffin-embedded Immunohistochemistry

Journal: Cancers

Article Title: Fibroblasts Promote Resistance to KRAS Silencing in Colorectal Cancer Cells

doi: 10.3390/cancers16142595

Figure Lengend Snippet: List of anti-human antibodies used for flow cytometry.

Article Snippet: CD104 , FITC , REA236 , 130-124-266 , Miltenyi Biotec.

Techniques: Cytometry

Journal: OncoTargets and therapy

Article Title: Integrated RNA Sequencing and Single-Cell Mass Cytometry Reveal a Novel Role of LncRNA HOXA-AS2 in Tumorigenesis and Stemness of Hepatocellular Carcinoma

doi: 10.2147/OTT.S272717

Figure Lengend Snippet: Purified Antibodies About the Stem-Like Cells Centric Panel

Article Snippet: CD104 , 173Yb , 58XB4 , Fluidigm , 3173008B.

Techniques: Purification

Journal: Scientific Reports

Article Title: Granzyme B is elevated in autoimmune blistering diseases and cleaves key anchoring proteins of the dermal-epidermal junction

doi: 10.1038/s41598-018-28070-0

Figure Lengend Snippet: GzmB Cleavage assays on dermal-epidermal junction proteins. ( a ) 4–20% SDS-PAGE western blot of GzmB-mediated cleavage of α6 integrin (α6 int) sub-unit with and without inhibitors serpin A3N (SA3N) and compound 20 (Com20). Black arrows indicate cleavage fragments and * indicates full-length proteins. At a concentration of 200 nM GzmB produces cleavage bands, and this cleavage is prevented by GzmB inhibitors. Full-length blot is presented in Supplemental Fig. S5. ( b ) 4–20% SDS-PAGE western blot of GzmB-mediated cleavage of β4 integrin sub-unit (β4 int) with and without inhibitors SA3N and Com20. Black arrows indicate cleavage fragments and * indicates full-length proteins. At a concentration of 200 nM GzmB produces cleavage bands, but its inhibition prevents the appearance of these bands. Full-length blot is presented in Supplemental Fig. . ( c ) 10% SDS-PAGE western blot of GzmB-mediated cleavage of collagen VII (coll VII) with and without inhibitors SA3N and Com20. Black arrows indicate cleavage fragments and * indicates full-length proteins. GzmB produces cleavage bands, and this cleavage is reduced by the addition of Com20 and abolished by SA3N. Full-length blot is presented in Supplemental Fig. . ( d ) 8% SDS-PAGE western blot of GzmB-mediated cleavage of collagen XVII (coll XVII) with and without inhibitors SA3N and Com20. Black arrow indicates cleavage fragment and * indicates full-length protein. GzmB produces a cleavage band, and this cleavage is abolished by the addition of Com20 or SA3N. Full-length blot is presented in Supplemental Fig. .

Article Snippet: Cleavage was detected by western blot using anti-human integrin α6 sub-unit (Cat # ab181551, ABCAM), anti-human integrin β4 sub-unit (Cat # MAB4060, R&D Systems, Minneapolis, MN), anti-collagen VII (Cat # ab93350, ABCAM), NC16A antibody directed against the NC16A domain of human collagen XVII and imaged using a Li-Cor Odissey FC (Li-Cor, Lincoln, NE).

Techniques: SDS Page, Western Blot, Concentration Assay, Inhibition

Journal: Scientific Reports

Article Title: Granzyme B is elevated in autoimmune blistering diseases and cleaves key anchoring proteins of the dermal-epidermal junction

doi: 10.1038/s41598-018-28070-0

Figure Lengend Snippet: Mass spectrometry analysis on GzmB-cleaved DEJ substrates. ( a ) Extracellular domain schematics for α6 integrin. GzmB mediated cleavage sites identified proteomically by ATOMs (red arrows) fall within the ligand-binding domains in the β-propeller region. GzmB, granzyme B; TM, transmembrane helix. ( b ) Extracellular domain schematics for β4 integrin. GzmB mediated cleavage sites identified proteomically by ATOMS (red arrows) falls within ligand-binding domains in the specificity-determining loop. ( c ) Extracellular domain schematics for collagen VII. GzmB mediated cleavage sites identified proteomically by ATOMS (red arrows) falls in the von Willebrand factor A and fibronectin type III-2 domains, which mediate collagen VII attachment to other dermal-epidermal junction components, such as laminins and collagen IV. TM, transmembrane helix; PSI, plexin-semaphorin-integrin; VWFA, von Willebrand factor A; CRR, cysteine-rich region; FNIII, fibronectin-like III domain; NC, non collagenous region; CMP, cartilage matrix protein; VWFA1/2, von Willebrand factor A 1/2; Pi, protein inhibitor.

Article Snippet: Cleavage was detected by western blot using anti-human integrin α6 sub-unit (Cat # ab181551, ABCAM), anti-human integrin β4 sub-unit (Cat # MAB4060, R&D Systems, Minneapolis, MN), anti-collagen VII (Cat # ab93350, ABCAM), NC16A antibody directed against the NC16A domain of human collagen XVII and imaged using a Li-Cor Odissey FC (Li-Cor, Lincoln, NE).

Techniques: Mass Spectrometry, Ligand Binding Assay

Journal: Scientific Reports

Article Title: Granzyme B is elevated in autoimmune blistering diseases and cleaves key anchoring proteins of the dermal-epidermal junction

doi: 10.1038/s41598-018-28070-0

Figure Lengend Snippet: Immunohistochemistry of DEJ components in healthy and diseased skin biopsies. α6 and β4 integrin, collagen VII, and collagen XVII immunostaining of healthy, bullous pemphigoid (BP), dermatitis herpetiformis (DH), and epidermolysis bullosa acquisita (EBA) biopsies. Blue arrowheads indicate intact protein in areas of dermo-epidermal adhesion, black arrowheads indicate weak or absent staining. Dotted lines indicate separation between the epidermis and the dermis. Scale bars represent 200 μm. α6 integrin is fragmented and scattered throughout the dermis in diseased skin, whereas in healthy biopsies this protein lines the DEJ. β4 integrin appears to be crucial for adhesion: in the bullous pemphigoid sample, a flap of dermis in the lower right corner is attached to the epidermis and shows strong β4 staining; this area is flanked by separated epidermis with faint β4 integrin staining. Scale bars represent 200 μm. Collagen VII lining is intact (blue arrowheads) in healthy skin, but weak or absent immunoreactivity was observed in diseased samples (black arrowheads). Collagen XVII lining is intact (blue arrowheads) in healthy skin, but weak or absent immunoreactivity was observed in diseased samples (black arrowheads).

Article Snippet: Cleavage was detected by western blot using anti-human integrin α6 sub-unit (Cat # ab181551, ABCAM), anti-human integrin β4 sub-unit (Cat # MAB4060, R&D Systems, Minneapolis, MN), anti-collagen VII (Cat # ab93350, ABCAM), NC16A antibody directed against the NC16A domain of human collagen XVII and imaged using a Li-Cor Odissey FC (Li-Cor, Lincoln, NE).

Techniques: Immunohistochemistry, Immunostaining, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: CD73 + Epithelial Progenitor Cells That Contribute to Homeostasis and Renewal Are Depleted in Eosinophilic Esophagitis

doi: 10.1016/j.jcmgh.2022.01.018

Figure Lengend Snippet: The CD73 + CD104 + population is depleted in the esophagus of active EoE patients and a murine model of EoE. ( A ) Immunofluorescence staining for CD73 and CD104 on biopsy specimens from non-EoE controls and patients with active EoE. Representative images are shown. Dotted line represents the basement membrane. Scale bars : 100 μm. ( B–E ) Biopsy specimens from non-EoE controls (n = 10), and patients with active (n = 10) or inactive EoE (n = 10) were dissociated for flow cytometry. The representative flow cytometry plots are shown in panel B . ( C ) The percentage of CD73 + CD104 + or CD73 - CD104 + cells in non-EoE controls (n = 10), and patients with active (n = 10) or inactive EoE (n = 10). ( D and E ) Pearson correlation coefficient analyses comparing the percentage of CD73 + CD104 + cells detected by flow cytometry and the number of eosinophils (eos/hpf) in ( D ) H&E-stained biopsy specimens or ( E ) CD45 + cells detected by flow cytometry. ( F and G ) After BALB/c mice were treated with the EoE protocol, their esophagi were resected for histologic sections or flow cytometry. ( F ) Histologic sections (H&E staining) of the esophagus from control (nontreated [NT]) or EoE protocol–treated mice. Arrowheads identify tissue-infiltrating eosinophils. ( G ) The percentage of CD73 + cells in esophageal epithelium of EoE mice (n = 3), compared with NT mice (n = 3). The data are shown as means ± SD. A 2-tailed Student t test or analysis of variance was performed for comparing 2 or multiple comparisons, respectively. ∗ P < .05.

Article Snippet: IHC of 3D organoids for CD73 and CD104 was performed by BOND RXm (Leica Biosystems, Nusslock, Germany) with anti-CD73 antibody (rabbit polyclonal, ab175396, 1:200; Abcam) or anti-CD104 monoclonal antibody (NBP2-37392, 1:1000, Novus Biologicals, Englewood, CA).

Techniques: Immunofluorescence, Staining, Membrane, Flow Cytometry, Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: CD73 + Epithelial Progenitor Cells That Contribute to Homeostasis and Renewal Are Depleted in Eosinophilic Esophagitis

doi: 10.1016/j.jcmgh.2022.01.018

Figure Lengend Snippet: Th2 cytokines shift the basaloid population from CD73 + to CD73 - in human esophageal 3D organoids. PDOs from non-EoE controls (n = 6) or patients with active EoE (n = 6), or immortalized esophageal epithelial (EPC2-hTERT) organoids were treated with IL4 (10 ng/mL) or IL13 (10 ng/mL) from day 7 to day 11. Nontreated (NT) organoids were used as control. ( A ) Representative flow cytometry plots showing the percentage of CD73 + CD104 + cells in PDOs from non-EoE controls or patients with active EoE, or EPC2-hTERT organoids, treated with IL4 or IL13. ( B ) The percentage of CD73 + CD104 + cells in each PDO treated with IL4 or IL13, paired with NT control. The data are shown as means ± SD. ∗∗∗ P < .001, ∗∗∗∗ P < .0001, 2-tailed Student t test. ( C ) The representative images of H&E staining, and IHC for CD73 or CD104 in EPC2-hTERT organoids treated with IL4 or IL13. Scale bars : 100 μm. ( D ) CD73 , CD104 , SOX2 , and IVL gene expression in EPC2-hTERT organoids treated with IL4 or IL13 were validated by qRT-PCR. The relative expression over NT is represented. The data are shown as means ± SD (n = 3). ∗ P < .05, ∗∗ P < .01, and ∗∗∗∗ P < .0001, analysis of variance.

Article Snippet: IHC of 3D organoids for CD73 and CD104 was performed by BOND RXm (Leica Biosystems, Nusslock, Germany) with anti-CD73 antibody (rabbit polyclonal, ab175396, 1:200; Abcam) or anti-CD104 monoclonal antibody (NBP2-37392, 1:1000, Novus Biologicals, Englewood, CA).

Techniques: Control, Flow Cytometry, Staining, Gene Expression, Quantitative RT-PCR, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: CD73 + Epithelial Progenitor Cells That Contribute to Homeostasis and Renewal Are Depleted in Eosinophilic Esophagitis

doi: 10.1016/j.jcmgh.2022.01.018

Figure Lengend Snippet: Single-cell RNA sequencing analysis of PDOs from 4 non-EoE patients. PDOs from 4 non-EoE patients were treated with IL13 (10 ng/mL) or vehicle (phosphate-buffered saline) from day 7 to day 11, and harvested and dissociated for single-cell RNA sequencing analysis on day 11. ( A ) Seurat’s Uniform Manifold Approximation and Projection (UMAP) visualization of 4 classified cell groups, basal-1, basal-2, suprabasal, and superficial, within PDOs from non-EoE controls grown with or without IL13 (10 ng/mL). ( B ) Average normalized gene expression values of MKI67 , CD73 , basal markers ( TP63 , NGFR ), and differentiated markers ( IVL , KRT13 , FLG ) in each cell cluster. Circle size reflects the percentage of cells with a nonzero expression level for the indicated genes. Color intensity reflects the average expression level across all cells within each cluster. ( C ) Expression z-scores for the top 8 up-regulated genes in each cluster. ( D ) The proportions of each cell cluster were compared in PDOs (n = 4) with IL13 or vehicle. ( E–G ) The normalized expression of ( E ) IVL and FLG , ( F ) SOX2 , and ( G ) CD73 in nontreated (NT) PDOs was compared with that in PDOs treated with IL13 for each cluster. ∗∗∗∗ P < .0001, Wilcoxon signed-rank test. ( H ) GO analysis of differentially expressed gene profiles of CD73 + CD104 + cells, compared with CD73 - CD104 + cells in NT PDOs.

Article Snippet: IHC of 3D organoids for CD73 and CD104 was performed by BOND RXm (Leica Biosystems, Nusslock, Germany) with anti-CD73 antibody (rabbit polyclonal, ab175396, 1:200; Abcam) or anti-CD104 monoclonal antibody (NBP2-37392, 1:1000, Novus Biologicals, Englewood, CA).

Techniques: RNA Sequencing, Saline, Gene Expression, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: CD73 + Epithelial Progenitor Cells That Contribute to Homeostasis and Renewal Are Depleted in Eosinophilic Esophagitis

doi: 10.1016/j.jcmgh.2022.01.018

Figure Lengend Snippet: CD73 + CD104 + cells represent a less-differentiated and slow-cycling cell population within the basal zone. ( A and B ) EPC2-hTERT organoids were harvested on day 11 and dissociated for FACS. ( A ) The cells in EPC2-hTERT organoids were sorted based on CD73 and CD104 expression. ( B ) CD73 , CD104 , IVL , and FLG relative gene expression in CD73 + CD104 + and CD73 + CD104 - populations was validated by qRT-PCR. ( C ) EPC2-hTERT cells in 2-dimensional culture (organoid day 0) and EPC2-hTERT organoid (day 7 and day 11) were harvested for flow cytometry and the representative flow cytometry plots for CD73 and CD104 expression are shown. ( D ) Representative IHC images for CD73 staining in EPC2-hTERT organoids harvested on day 5, day 7, day 9, and day 11. Scale bars : 100 μm. ( E–G ) EPC2-hTERT organoids were harvested on day 11 after incubation in culture medium with EdU for 2 hours, and dissociated for flow cytometry. ( E ) A representative FACS dot plot of cells from EPC2-hTERT organoids subjected to EdU, with the frequencies of each population according to CD73 and CD104 expression, is shown. ( F ) Representative histograms of EdU staining in each cell population. The percentage of EdU + cells in each population is shown. ( G ) Percentages of EdU + cells in each population. All data are shown as means ± SD (n = 3). A 2-tailed Student t test or analysis of variance were performed for comparing 2 or multiple comparisons, respectively. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, and ∗∗∗∗ P < .0001.

Article Snippet: IHC of 3D organoids for CD73 and CD104 was performed by BOND RXm (Leica Biosystems, Nusslock, Germany) with anti-CD73 antibody (rabbit polyclonal, ab175396, 1:200; Abcam) or anti-CD104 monoclonal antibody (NBP2-37392, 1:1000, Novus Biologicals, Englewood, CA).

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Flow Cytometry, Staining, Incubation

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: CD73 + Epithelial Progenitor Cells That Contribute to Homeostasis and Renewal Are Depleted in Eosinophilic Esophagitis

doi: 10.1016/j.jcmgh.2022.01.018

Figure Lengend Snippet: CD73 + cells mediate epithelial renewal in human esophageal epithelium. ( A–D ) PDOs or EPC2-hTERT organoids were harvested on day 11 (1° organoids) and dissociated for FACS. The cells were sorted based on CD73 and CD104 expression and resuspended in Matrigel for organoid establishment and growth. OFR and organoid size (mean area) of 2° organoids were ( B and C ) quantified on a Celigo image cytometer and ( D ) harvested on day 11 and again dissociated for flow cytometry. ( A ) Representative phase-contrast images of organoids formed by corresponding cell populations. ( B ) OFR and ( C ) mean size were compared among organoids from the 3 populations. ( D ) The scheme of this experiment and the representative flow cytometry plots of 2° organoids are shown. The percentage of CD73 + cells in EPC2-hTERT were compared. The data are shown as means ± SD (n = 3). ( E–H ) EPC2-hTERT cells were treated for 48 hours with CD73 siRNA or nonsilencing (scramble) siRNA in 2D culture, followed by seeding into organoid cultures. ( E ) CD73 silencing efficiency was evaluated by flow cytometry. ( F ) Representative phase-contrast images of organoids formed by corresponding cells. ( G ) OFR and ( H ) mean size of organoids from EPC2-hTERT cells transfected with CD73 or scramble siRNA. The data are shown as means ± SD (n = 10). Two-tailed Student t test or analysis of variance were performed for single or multiple comparisons, respectively. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, and ∗∗∗∗ P < .0001.

Article Snippet: IHC of 3D organoids for CD73 and CD104 was performed by BOND RXm (Leica Biosystems, Nusslock, Germany) with anti-CD73 antibody (rabbit polyclonal, ab175396, 1:200; Abcam) or anti-CD104 monoclonal antibody (NBP2-37392, 1:1000, Novus Biologicals, Englewood, CA).

Techniques: Expressing, Cytometry, Flow Cytometry, Transfection, Two Tailed Test

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: CD73 + Epithelial Progenitor Cells That Contribute to Homeostasis and Renewal Are Depleted in Eosinophilic Esophagitis

doi: 10.1016/j.jcmgh.2022.01.018

Figure Lengend Snippet: The therapeutic potential of STAT6 inhibition in EoE. ( A ) IHC staining for phosphorylated STAT6 was performed, with representative images shown. Scale bars : 100 μm. ( B ) The positive rate of phosphorylated STAT6 in esophageal epithelium are compared. The data are shown as means ± SD (n = 5). ( C ) Relative expression of CD73 gene in EPC2-hTERT organoids treated with STAT6 inhibitor (AS151749, 400 nmol/L), IL4 (10 ng/mL), or IL13 (10 ng/mL). ( D ) The frequencies of CD73 + CD104 + cells in organoids treated with AS151749 or dimethyl sulfoxide (DMSO) under the presence of Th2 cytokines. STAT6 silencing efficiency was evaluated by ( E ) qRT-PCR and ( F ) Western blot. ( G ) CD73 gene expression in EPC2-hTERT organoids treated with STAT6 siRNA or nonsilencing (scramble) siRNA with or without IL13. The data are shown as means ± SD (n = 3). Analysis of variance was performed with multiple comparisons. ∗∗ P < .01, ∗∗∗∗ P < .0001.

Article Snippet: IHC of 3D organoids for CD73 and CD104 was performed by BOND RXm (Leica Biosystems, Nusslock, Germany) with anti-CD73 antibody (rabbit polyclonal, ab175396, 1:200; Abcam) or anti-CD104 monoclonal antibody (NBP2-37392, 1:1000, Novus Biologicals, Englewood, CA).

Techniques: Inhibition, Immunohistochemistry, Expressing, Quantitative RT-PCR, Western Blot, Gene Expression

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: CD73 + Epithelial Progenitor Cells That Contribute to Homeostasis and Renewal Are Depleted in Eosinophilic Esophagitis

doi: 10.1016/j.jcmgh.2022.01.018

Figure Lengend Snippet: Omeprazole treatment prevents Th2-mediated depletion of CD73 + CD104 + population. ( A ) EPC2-hTERT organoids were treated with omeprazole (50 μmol/L), IL4 (10 ng/mL), or IL13 (10 ng/mL). The frequencies of CD73 + CD104 + cells in organoids are shown as means ± SD (n = 3). ( B and C ) The treated EPC2-hTERT organoids were harvested on day 11 and dissociated into single cells, followed by seeding in Matrigel to assess organoid formation capacity. ( B ) OFR was evaluated manually on day 11. ( C ) Representative phase-contrast images of organoids from corresponding groups. The data are shown as means ± SD (n = 6). Analysis of variance was performed with multiple comparisons. ∗∗∗∗ P < .0001. DMSO, dimethyl sulfoxide.

Article Snippet: IHC of 3D organoids for CD73 and CD104 was performed by BOND RXm (Leica Biosystems, Nusslock, Germany) with anti-CD73 antibody (rabbit polyclonal, ab175396, 1:200; Abcam) or anti-CD104 monoclonal antibody (NBP2-37392, 1:1000, Novus Biologicals, Englewood, CA).

Techniques:

Journal: Marine Drugs

Article Title: Characterisation of Elevenin-Vc1 from the Venom of Conus victoriae : A Structural Analogue of α-Conotoxins

doi: 10.3390/md21020081

Figure Lengend Snippet: Two-dimensional [ 15 N– 1 H]-HSQC spectrum of elevenin-Vc1 showing backbone amide resonance assignments. The spectrum was acquired at 303 K and pH 4 on a Bruker Avance III 600 MHz spectrometer.

Article Snippet: The human muscle nAChR clones (α1, β1, δ, and ε) were purchased from Integrated DNA Technologies (Coralville, IA, USA), the human (h) α3, α9, α10, β2, and β4 clones were purchased from OriGene (Rockville, MD, USA), and all were subsequently inserted into the pT7TS vector.

Techniques:

Journal: Marine Drugs

Article Title: Characterisation of Elevenin-Vc1 from the Venom of Conus victoriae : A Structural Analogue of α-Conotoxins

doi: 10.3390/md21020081

Figure Lengend Snippet: ( A ) Region of 600 MHz NOESY spectrum of elevenin-Vc1 showing NOEs diagnostic of the Ala11-Pro12 peptide bond conformation. Strong NOEs were observed between Ala11 Hα ( i ) and Pro12 Hδ protons ( i + 1), which are ~2.2 Å apart, indicating the trans conformation of the Ala11-Pro12 bond ( B ). In addition, a weak NOE (⁎) is observed between Ala11 Hα and Pro12 Hα protons; this may be because these protons are within 5 Å (4.4 Å). ( C ) 2D [ 13 C– 1 H]-HSQC spectrum showing Pro12 Cα and Cβ NMR assignments. The small Cβ and Cγ chemical shift difference of 4.3 ppm (<5 ppm for trans and ~10 ppm for cis ) further confirms the trans geometry of the Ala11-Pro12 peptide bond.

Article Snippet: The human muscle nAChR clones (α1, β1, δ, and ε) were purchased from Integrated DNA Technologies (Coralville, IA, USA), the human (h) α3, α9, α10, β2, and β4 clones were purchased from OriGene (Rockville, MD, USA), and all were subsequently inserted into the pT7TS vector.

Techniques: Diagnostic Assay