Journal: Technology in Cancer Research & Treatment

Article Title: B7-H3 and CD39 Co-Localization in Gastric Cancer: A Potential Prognostic Biomarker and Potential Dual-Target for Immunotherapy

doi: 10.1177/15330338251380957

Figure Lengend Snippet: Expression Patterns of B7-H3 and CD39 in Gastric Precancerous Lesions and Gastric Cancer Tissues. (A) Representative IHC Expression Patterns of B7-H3 and CD39 Across Different Pathological Stages (CSG, CAG, LGIN, HGIN, and GC). (B) Quantitative Analysis of Positive Expression Areas of CD39. (C) Quantitative Analysis of Positive Expression Areas of B7-H3. (D) Representative Multiplex IHC Expression Patterns of B7-H3 and CD39 Across Different Pathological Stages (CSG, CAG, LGIN, HGIN, and GC). (E) Quantitative Analysis of Multiplex IHC Expression of B7-H3 and CD39. (F) Representative Co-Localization Expression of B7-H3 and CD39 in GC. IHC Scale Bars: 50 µm; mIHC Scale Bars: 100 µm.

Article Snippet: The following primary antibodies were used according to the manufacturer's instructions: mouse anti-human B7-H3 monoclonal antibody, 1/200 dilution (Cat No: 66 481-1-Ig, Proteintech, China), rabbit anti-human CD39 polyclonal antibody, 1/1000 dilution (Cat No: 14211-1-AP, Proteintech, China) and mouse anti-human CD8 monoclonal antibody, 1/10 000 dilution (Cat No: 66868-1-Ig, Proteintech, China).

Techniques: Expressing, Multiplex Assay

Journal: Technology in Cancer Research & Treatment

Article Title: B7-H3 and CD39 Co-Localization in Gastric Cancer: A Potential Prognostic Biomarker and Potential Dual-Target for Immunotherapy

doi: 10.1177/15330338251380957

Figure Lengend Snippet: Co-Localization of B7-H3 and CD39 in Gastric Cancer Cells Indicates Poor Prognosis. (A, B) Representative Immunohistochemical Images Showing Low and High Expression of B7-H3 (A) and CD39 (B) in Gastric Cancer (GC) Specimens. (C) Correlation Analysis of B7-H3 and CD39 Expression. (F, G, H, I) Kaplan-Meier Survival Curves for Overall Survival of GC Patients Based on the Expression Status of B7-H3 (F), CD39 (G), Dual High Expression of B7-H3 and CD39 (H), and Co-Localized Expression Status of B7-H3-CD39 (I). Scale Bars: 100 µm.

Article Snippet: The following primary antibodies were used according to the manufacturer's instructions: mouse anti-human B7-H3 monoclonal antibody, 1/200 dilution (Cat No: 66 481-1-Ig, Proteintech, China), rabbit anti-human CD39 polyclonal antibody, 1/1000 dilution (Cat No: 14211-1-AP, Proteintech, China) and mouse anti-human CD8 monoclonal antibody, 1/10 000 dilution (Cat No: 66868-1-Ig, Proteintech, China).

Techniques: Immunohistochemical staining, Expressing

Journal: Technology in Cancer Research & Treatment

Article Title: B7-H3 and CD39 Co-Localization in Gastric Cancer: A Potential Prognostic Biomarker and Potential Dual-Target for Immunotherapy

doi: 10.1177/15330338251380957

Figure Lengend Snippet: Absence of Correlation Between Co-localization of B7-H3 and CD39 Expression and CD8 + T Cell Infiltration in Gastric Cancer. (A, B) Representative Immunohistochemical Images Showing the Expression of B7-H3 with CD8 (A) or CD39 with CD8 (B) in the Same Patient. (C) Correlation Between B7-H3 Expression and CD8 Expression, and Between CD39 Expression and CD8 Expression. (D, E) Representative Multiplex Immunohistochemistry Images of B7-H3, CD39, and CD8 (D), and Correlation Analysis Between the Co-Localization Expression Score of B7-H3 and CD39 and the Extent of CD8 Infiltration (E). (F, G) Representative Multiplex Immunohistochemistry Images of CD39 and CD8, with Arrows Indicating CD39 + CD8 + T Cells (F), and the Proportion of CD39 + CD8 + T Cells within the CD8 + T Cell Population (G). Scale Bars: 100 µm.

Article Snippet: The following primary antibodies were used according to the manufacturer's instructions: mouse anti-human B7-H3 monoclonal antibody, 1/200 dilution (Cat No: 66 481-1-Ig, Proteintech, China), rabbit anti-human CD39 polyclonal antibody, 1/1000 dilution (Cat No: 14211-1-AP, Proteintech, China) and mouse anti-human CD8 monoclonal antibody, 1/10 000 dilution (Cat No: 66868-1-Ig, Proteintech, China).

Techniques: Expressing, Immunohistochemical staining, Multiplex Assay, Immunohistochemistry

Journal: Technology in Cancer Research & Treatment

Article Title: B7-H3 and CD39 Co-Localization in Gastric Cancer: A Potential Prognostic Biomarker and Potential Dual-Target for Immunotherapy

doi: 10.1177/15330338251380957

Figure Lengend Snippet: Kaplan-Meier Survival Curves for Gastric Cancer Patients Based on Specific Expression Statuses and Immune Cell Infiltration Levels. (A, B, C, D) Kaplan-Meier Survival Curves for GC Patients Stratified by B7-H3 High CD8 Low (A) Expression Status 、 CD39 High CD8 Low (B) Expression Status 、 B7-H3-CD39 Both high CD8 Low Expression Status (C) and B7-H3-CD39 Co-localization CD8 low Expression Status (D). (E, F) Kaplan-Meier Survival Curves for GC Patients Based on CD8 + T Cell Infiltration Levels (E) and CD39 + CD8 + T Cell Infiltration Status (F).

Article Snippet: The following primary antibodies were used according to the manufacturer's instructions: mouse anti-human B7-H3 monoclonal antibody, 1/200 dilution (Cat No: 66 481-1-Ig, Proteintech, China), rabbit anti-human CD39 polyclonal antibody, 1/1000 dilution (Cat No: 14211-1-AP, Proteintech, China) and mouse anti-human CD8 monoclonal antibody, 1/10 000 dilution (Cat No: 66868-1-Ig, Proteintech, China).

Techniques: Expressing

Journal: Frontiers in immunology

Article Title: Immunometabolic interference between cancer and COVID-19.

doi: 10.3389/fimmu.2023.1168455

Figure Lengend Snippet: FIGURE 9 Flow cytometry analysis of NAMPT and NAD-dependent enzymes in the different patient cohorts. Box-and-whisker plots representing FACS quantification of (A) NAMPT, (B) pSIRT1, (C) CD38, (D) CD39 and (E) PARP expression in the indicated immune cell populations (CD3+ T cells, CD19+

Article Snippet: Cells were incubated with the following antihuman antibodies for 20 min at 4°C: CD33 PerCP Cy5.5 (WM53) (0.6:100, Biolegend cat No. 303402), CD14 BV650 (M5E2) (1.25:100, Biolegend cat. No. 301835), CD16 BV711 (3G8) (0.3:100, Biolegend cat. No. 302044), HLA-DR BV421 (L243) (1.25:100, Biolegend Cat No. 307636), CD15 BV786 (HI98) (1.25:100, BD Biosciences cat. No. 563838), CD11c BUV661 (B-ly6) (0.6:100, BD Biosciences cat. No. 612968), CD123 BUV395 (7G3) (0.6:100, BD Biosciences cat. No. 564195), CD38 PE-Cy7 (HIT2) (1.25:100, Invitrogen cat. No. 25- 0389-42), CD39 PE (MZ18-23C8) (0.6:100, MACSMiltenyi Biotec Inc. cat. No.130-118-668), CD39 FITC (MZ18-23C8) (1.25:100, MACS Miltenyi Biotec Inc. cat. No. 130-125-113), CD3 BV650 (OKT3) (1.25:100, Biolegend cat. No.317324), CD4 BV570 (OKT4) (2.5:100, Frontiers in Immunology 03 Biolegend cat. No. 317445), CD8 BV786 (RPA-T8) (0.6:100, BD Biosciences cat. No. 557085), CD19 AF-700 (HIB19) (2.5:100, BD Biosciences cat. No. 561031), CD45RA APC-Cy7 (HI100) (1.25:100, BD Biosciences cat. No. 560674), CD45RO BUV395 (UCHL1) (1.25:100, BD Biosciences 564292), CD27 PE-Cy5 (O323) (1.25:100, Invitrogen cat. No. 15295964), CD16 BV605 (3G8) (0.6:100, Biolegend 302040), CD3 BUV496 (UCHT1) (1.25:100, BD Biosciences cat. No. 612940), CD66b PE-Cy7(G10F5) (0.6:100 Biolegend cat. No. 305116) and IgD FITC (IA6-2) (1.25:100, Invitrogen cat. No. 11-9868-42).

Techniques: Flow Cytometry, Whisker Assay, 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: CD39 , REA739 , 50 , 130-110-650 , PE , Miltenyi Biotec.

Techniques: Imaging

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: Cellular neighborhood analysis of PD1 high/low T cells in the tumor margin and core. (A–D) Topology of PD1 high (left) and PD1 low (right) T cells and their cellular neighborhood within a 5 µm range. (A, B) represent tumor margin and (C, D) show tumor core areas. Cell types showing different distribution patterns around PD1 high and PD1 low T cells (mDCs, M1-like M, M2-like M, MDSCs, Fibroblasts, vessels, tumor cells) are highlighted by arrowheads. (E, F) Quantification of cells in a 5 µm range around of PD1 high/low T cells for tumor margin and tumor core, (E) represents immune cells and (F) stroma/tumor cells. (G) Violin plots for expression levels of eight immune-modulating markers (CD112, CD155, CD276, CD39, CD73, IDO, PD-L1, and VISTA) for the most important immune and tumor cells around PD1 high/low T cells in the tumor core area. Violin plots for tumor margin are shown in Supplementary Figure S5E . Depicted markers and annotated cell types as indicated by the color code. ROI sizes: Tumor margin (ROI15) and tumor core (ROI16): 975 x 769 µm.

Article Snippet: CD39 , REA739 , 50 , 130-110-650 , PE , Miltenyi Biotec.

Techniques: Expressing

Journal: Science Advances

Article Title: Dysregulation of ectonucleotidase-mediated extracellular adenosine during postmenopausal bone loss

doi: 10.1126/sciadv.aax1387

Figure Lengend Snippet: ( A to K ) Characterization of healthy (sham) and OVX animals 4 weeks after ovariectomy. (A) Immunofluorescence staining of CD73 (green) and (B) CD39 (red) in vertebrae of OVX animals. Nuclear staining (blue). Scale bars, 100 μm. Inset shows magnified image of bone surface. Yellow arrowheads indicate cells positive for CD73 or CD39 on bone surface. Scale bars, 50 μm. (C) Flow cytometric analysis of CD73 and CD39 membrane expression of hematopoietic cells from mouse BM cells 4 weeks after ovariectomy (OVX) and healthy controls. (D) Percentage and median fluorescence intensity of hematopoietic cells expressing CD73. (E) Percentage and median fluorescence intensity of hematopoietic cells expressing CD39. (F) Flow cytometric analysis of CD73 and CD39 membrane expression of nonhematopoietic cells from mouse BM cells 4 weeks after ovariectomy and healthy controls. (G) Percentage and median fluorescence intensity of nonhematopoietic cells expressing CD73. (H) Percentage and median fluorescence intensity of nonhematopoietic cells expressing CD39. (I) CD73 gene expression and (J) CD39 gene expression of cells from bone chips. (K) Extracellular adenosine concentration in BM plasma of sham and OVX animals. n = 5. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Sections were then incubated with primary antibody against CD39 (5 μg/ml; AF4398, R&D Systems), CD73 (5 μg/ml; AF4488, R&D Systems), and A2BR (1:200; MBS8207549, MyBioSource, San Diego, CA) in diluent solution (1%, w/v) and normal donkey serum (1%, v/v) in TBS overnight at 4°C.

Techniques: Immunofluorescence, Staining, Membrane, Expressing, Fluorescence, Gene Expression, Concentration Assay, Clinical Proteomics

Journal: Science Advances

Article Title: Dysregulation of ectonucleotidase-mediated extracellular adenosine during postmenopausal bone loss

doi: 10.1126/sciadv.aax1387

Figure Lengend Snippet: ( A ) Flow cytometric analyses and ( B ) quantification of CD73 and CD39 in osteoprogenitors in the absence or presence of E2 (100 nM) for 3 days. ( C to E ) Single (ESR1 or ESR2) or dual (ESR1 and ESR2) ER knockdown (KD) by siRNA in primary mouse osteoprogenitors and analyzed after 3 days. (C) Flow cytometric analyses of CD73 and CD39 after single knockdown (ESR1 or ESR2) and dual knockdown (ESR1 and ESR2). (D) Percentage of double-positive (CD73/CD39) cells in single knockdown and dual knockdown cells. (E) In vitro adenosine levels normalized by cell number in single knockdown and dual knockdown cells. Control (scrambled) siRNA concentration for single knockdown and dual knockdown are 5 and 10 nM, respectively. n = 5. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Sections were then incubated with primary antibody against CD39 (5 μg/ml; AF4398, R&D Systems), CD73 (5 μg/ml; AF4488, R&D Systems), and A2BR (1:200; MBS8207549, MyBioSource, San Diego, CA) in diluent solution (1%, w/v) and normal donkey serum (1%, v/v) in TBS overnight at 4°C.

Techniques: Knockdown, In Vitro, Control, Concentration Assay

Journal: Science Advances

Article Title: Dysregulation of ectonucleotidase-mediated extracellular adenosine during postmenopausal bone loss

doi: 10.1126/sciadv.aax1387

Figure Lengend Snippet: ( A ) Flow cytometric analyses and ( B ) quantification of CD73 and CD39 in primary mouse mononuclear cells undergoing osteoclast differentiation in the absence or presence of E2 (100 nM) for 3 days. ( C to E ) Single (ESR1 or ESR2) or dual (ESR1 and ESR2) ER knockdown by siRNA during macrophage differentiation for 3 days and subsequent osteoclast differentiation for 6 days. (C) Flow cytometric analyses of CD73 and CD39 after single knockdown (ESR1 or ESR2) and dual knockdown (ESR1 and ESR2). (D) Percentage of double-positive (CD73/CD39) cells in single knockdown and dual knockdown cells. (E) In vitro adenosine levels normalized by cell number in single knockdown and dual knockdown cells. Control (scrambled) siRNA concentration for single knockdown and dual knockdown are 5 and 10 nM, respectively. n = 4. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Sections were then incubated with primary antibody against CD39 (5 μg/ml; AF4398, R&D Systems), CD73 (5 μg/ml; AF4488, R&D Systems), and A2BR (1:200; MBS8207549, MyBioSource, San Diego, CA) in diluent solution (1%, w/v) and normal donkey serum (1%, v/v) in TBS overnight at 4°C.

Techniques: Knockdown, In Vitro, Control, Concentration Assay

Journal: Cell Death & Disease

Article Title: P2X7 a new therapeutic target to block vesicle-dependent metastasis in colon carcinoma: Role of the A2A/CD39/CD73 axis

doi: 10.1038/s41419-025-07897-2

Figure Lengend Snippet: A CT26 CRC cells were stained with PKH26GL (red) and quinacrine (green) fluorescent dyes. Images were acquired using confocal microscopy at time 0 and after 5 min following P2X7 activation with 300 µM BzATP and are extrapolated from a 30-min time course (see supplementary videos and ). B CT26 cells were pre-treated with P2X7 antagonist AZ10606120 (5 µM) for 10 min before application of BzATP. C Number of vesicles released in 30 min from CT26 cells in PBS vehicle (PBS S-VS), following stimulation with P2X7 agonist ATP (P2X7-VS) or 10 min pretreatment with P2X7 antagonist AZ10606120 followed by stimulation with 3 mM ATP (AZ-VS) ( n = 5). D Size of PBS S-VS, P2X7-VS, and AZ-VS ( n = 5). E Number of vesicles released in 30 min from CT26 cells in DMSO vehicle (DMSO S-VS), following stimulation with P2X7 agonist ATP (P2X7-VS) or 10 min pretreatment with P2X7 antagonist A740003 followed by stimulation with 3 mM ATP (A74-VS) ( n = 7). F Size of DMSO- S-VS, P2X7-VS, and A74-VS ( n = 7). G Western blot for GM130, Alix, P2X7, CD39, CD73, and A2A in CT26 cells, S-VS and P2X7-VS. Pericellular ATP was measured with the pmeLUC probe expressed on the cell surface of untreated CT26 cells or after 5 min of exposure to PBS vehicle, S-VS, and P2X7-VS ( n = 4). H Quantification of luminescence changes was expressed as a fold increase on time 0. I Representative images of photon emissions. Changes in ATP J concentration increase on time 0 in the supernatants of CT26 cells, untreated or treated with PBS vehicle, S-VS, or P2X7-VS, measured with a luciferin/luciferase assay ( n = 3). Changes in adenosine K concentration increase on time 0 in the supernatants of CT26 cells untreated or treated with PBS vehicle, S-VS, P2X7-VS, or P2X7-VS plus 5uM CD73 inhibitor AB680 ( n = 5). * p < 0.05, ** p < 0.001, *** p < 0,0001, **** p < 0.00001.

Article Snippet: Tissue slides from the mouse lungs and rat colons were analyzed for P2X7, CD39, CD73, and A2A expression using the following primary antibodies: P2X7 1:100 (P8232, Sigma-Aldrich), CD39 1:100 (NBP2-67230, Novus Biologicals, Minneapolis, Minnesota, USA), CD73 1:500 (MAB5795, R&D Systems, Minneapolis, Minnesota, USA), and A2A 1:100 (SC32261, Santa Cruz Biotechnology).

Techniques: Staining, Confocal Microscopy, Activation Assay, Western Blot, Concentration Assay, Luciferase

Journal: Cell Death & Disease

Article Title: P2X7 a new therapeutic target to block vesicle-dependent metastasis in colon carcinoma: Role of the A2A/CD39/CD73 axis

doi: 10.1038/s41419-025-07897-2

Figure Lengend Snippet: The mRNA expression of P2X7A A , B , P2X7B C , D , CD39 E , F , CD73 G , H , and A2A I , J was evaluated in the cDNAs of 158 patients with CRC subdivided into stage I ( n = 24), stage II ( n = 50), stage III ( n = 52), and stage IV ( n = 32) which comprised 11 samples derived from metastases in organs other than the colon. K Spearman’s correlation coefficient among P2X7A, P2X7B, CD39, CD73 , and A2A was evaluated in CRC metastatic patients. L Spearman’s correlation coefficient was evaluated between P2X7 and A2A expression in colon adenocarcinoma samples obtained from the Cancer Genome Atlas database. * p < 0.05, ** p < 0.01, *** p < 0.001. **** p < 0.0001.

Article Snippet: Tissue slides from the mouse lungs and rat colons were analyzed for P2X7, CD39, CD73, and A2A expression using the following primary antibodies: P2X7 1:100 (P8232, Sigma-Aldrich), CD39 1:100 (NBP2-67230, Novus Biologicals, Minneapolis, Minnesota, USA), CD73 1:500 (MAB5795, R&D Systems, Minneapolis, Minnesota, USA), and A2A 1:100 (SC32261, Santa Cruz Biotechnology).

Techniques: Expressing, Derivative Assay

Journal: Cell Death & Disease

Article Title: P2X7 a new therapeutic target to block vesicle-dependent metastasis in colon carcinoma: Role of the A2A/CD39/CD73 axis

doi: 10.1038/s41419-025-07897-2

Figure Lengend Snippet: mRNA expression of A P2X7A , B P2X7B , C A2A , D CD39 , and E CD73 in CRC patients subdivided into APC WT and APC mutated groups ( n = 6). Percentage of cells positive for P2X7 F and A2A G in the colons of WT and PIRC rats and PIRC tumors ( n = 4). Representative images of immunohistochemical staining for P2X7 and A2A in the colon of WT 1-year rats H, K and in the normal colon I, L and the tumor mass J, M of 1-year PIRC rats. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Tissue slides from the mouse lungs and rat colons were analyzed for P2X7, CD39, CD73, and A2A expression using the following primary antibodies: P2X7 1:100 (P8232, Sigma-Aldrich), CD39 1:100 (NBP2-67230, Novus Biologicals, Minneapolis, Minnesota, USA), CD73 1:500 (MAB5795, R&D Systems, Minneapolis, Minnesota, USA), and A2A 1:100 (SC32261, Santa Cruz Biotechnology).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Nature Communications

Article Title: CD39 polymorphism enables lung thrombosis in sickle cell disease

doi: 10.1038/s41467-026-68396-2

Figure Lengend Snippet: a Experimental scheme for ( b – f , j ). EVs Extracellular vesicles, SEC Size exclusion chromatography, NTA Nanoparticle tracking analysis. Created in BioRender. Brzoska, T https://BioRender.com/t6bmaj7 . b NTA plot showing concentration vs size distribution of EVs isolated from a control and an SCD mouse plasma. c EV concentration in plasma of SCD (n = 7) and control (n = 7) mice. d Western blot micrograph and e the densitometry analysis (arbitrary units) of CD39 protein expression in control (n = 5) and SCD (n = 5) mice EVs. Ponceau-S, loading control. f ADPase activity in control (n = 5) and SCD (n = 5) mice EVs ± incubation with CD39 inhibitor (500 µM ARL67156 ). g Experimental scheme for ( h , i ). Platelet-rich plasma, PRP. Created in BioRender. Brzoska, T. (2025) https://BioRender.com/vq6jifk . h In vitro platelet aggregation kinetics in a control mouse PRP sample following the addition of ADP (black), ADP + control mouse EVs (red), ADP + SCD mouse EVs (blue), and ADP + SCD mouse EVs + POM-1 (green). i Area under the curve (AUC) in four groups shown in ( h ). N = 4 per group. j Imaging flow cytometry images of CD39 + /CD31 + /CD144 + (row #1) or CD39 + /CD31 + /CD106 + (row #2) EVs isolated from SCD mice plasma. Bottom row- isotype control Ab stained EVs. Scale bar, 5 µm. Data representative of 3 independent experiments. k Experimental scheme for ( l – n ). In vitro cultured human lung microvascular endothelial cells (HMVECs-L) ± incubation with 20 µM hemin and EVs isolated from cell culture supernatant. Created in BioRender. Brzoska, T https://BioRender.com/yfnjlra . l NTA plot showing concentration vs size distribution of HMVECs-L EVs. m EV concentration in the supernatant of HMVECs-L incubated with (n = 6 independent experiments) or without (n = 6 independent experiments) hemin. n ADPase activity in EVs isolated from the supernatant of HMVECs-L incubated with (n = 4 independent experiments) or without (n = 4 independent experiments) hemin ±500 µM ARL67156 . Means were compared using unpaired two-tailed Student’s t test. Data represent mean ± SEM. Exact P values shown in the graphs.

Article Snippet: For each treatment group, 20 μl of EVs suspension was diluted with 80 μl of sterile PBS and incubated in the dark for 15 min (4 °C) with Alexa Fluor 647 anti-human CD39 antibody (clone: 498403; R&D Systems cat# FAB4397R) and Phycoerythrin (PE) anti-human CD31 antibody (clone: WM59, BD Pharmingen cat# 560983) for in situ staining of CD39 and CD31, respectively.

Techniques: Size-exclusion Chromatography, Concentration Assay, Isolation, Control, Clinical Proteomics, Western Blot, Expressing, Activity Assay, Incubation, In Vitro, Imaging, Flow Cytometry, Staining, Cell Culture, Two Tailed Test

Journal: Nature Communications

Article Title: CD39 polymorphism enables lung thrombosis in sickle cell disease

doi: 10.1038/s41467-026-68396-2

Figure Lengend Snippet: a Number of SCD patients without (-PT) vs with (+PT) medical history of pulmonary thrombosis in Walk-PHASST registry. Created in BioRender. Brzoska, T https://BioRender.com/aovqt1d . Frequency of b rs3176891 GG genotype and ( c ) rs3176891G allele among -PT vs + PT SCD patients in ( a ). Data compared using four-fold table analysis with two-tailed χ2 test. d Odds Ratio for association of rs3176891G allele with risk of pulmonary thrombosis in SCD (n = 437) and non-SCD (n = 1891) humans. Data represent point estimate of Odds Ratio ±95% CI. e Number of non-SCD humans of African ancestry without (-PT) vs with ( + PT) medical history of pulmonary thrombosis in TOPMed database. Created in BioRender. Brzoska, T. (2025) https://BioRender.com/aovqt1d . f Frequency of rs3176891G allele among non-SCD humans in ( e ). g Experimental scheme for ( h – n ): blood from SCD patients with AA or AG or GG genotype of rs3176891 processed to generate platelet-rich plasma (PRP) and platelet-free plasma (PFP). Created in BioRender. Brzoska, T https://BioRender.com/ d8skzoa. h ADPase activity in EVs of SCD patients with AA (n = 4) vs AG/GG (n = 10) genotype. ADPase activity in EVs of SCD patients with i AA (n = 4) and j AG/GG (n = 10) genotype ± incubation with 500 µM ARL67156 . k Imaging flow cytometry images showing CD39 + /CD31 + EVs in the plasma of an SCD patient with AA genotype. Scale bar 5 µm. l Concentration of CD39 + /CD31 + EVs in PFP of SCD patients with AA (n = 3) vs AG/GG (n = 4) genotype. In vitro platelet aggregation kinetics shown as the percent increase in light transmission in PRP of SCD patients with AA (black), AG (blue) and GG (red) genotypes, following the addition of m 1 µM ADP or n 3 µg/ml collagen. Data from more patients are shown as Supplementary Fig. . Data compared using unpaired two-tailed Student’s t test in ( h , l ) (mean ± SEM), paired two-tailed Student’s t test in ( i ), and two-tailed Wilcoxon matched-pairs signed rank test in ( j ). Exact P values shown in the graphs.

Article Snippet: For each treatment group, 20 μl of EVs suspension was diluted with 80 μl of sterile PBS and incubated in the dark for 15 min (4 °C) with Alexa Fluor 647 anti-human CD39 antibody (clone: 498403; R&D Systems cat# FAB4397R) and Phycoerythrin (PE) anti-human CD31 antibody (clone: WM59, BD Pharmingen cat# 560983) for in situ staining of CD39 and CD31, respectively.

Techniques: Two Tailed Test, Clinical Proteomics, Activity Assay, Incubation, Imaging, Flow Cytometry, Concentration Assay, In Vitro, Transmission Assay

Journal: Nature Communications

Article Title: CD39 polymorphism enables lung thrombosis in sickle cell disease

doi: 10.1038/s41467-026-68396-2

Figure Lengend Snippet: Although ADP released during acute intravascular hemolysis can trigger in situ pulmonary thrombosis by stimulating platelet-purinergic P2Y1 and P2Y12 receptors, the sterile inflammatory milieu in SCD also promotes the generation of endothelium-derived CD39 + EVs that phosphohydrolize ADP to prevent pulmonary thrombosis. However, ENTPD1 rs3176891G allele is associated with impaired generation of CD39 + EVs, thus increasing the risk of pulmonary thrombosis in some SCD patients. Created in BioRender. Brzoska, T. (2025) https://BioRender.com/x4rvhil .

Article Snippet: For each treatment group, 20 μl of EVs suspension was diluted with 80 μl of sterile PBS and incubated in the dark for 15 min (4 °C) with Alexa Fluor 647 anti-human CD39 antibody (clone: 498403; R&D Systems cat# FAB4397R) and Phycoerythrin (PE) anti-human CD31 antibody (clone: WM59, BD Pharmingen cat# 560983) for in situ staining of CD39 and CD31, respectively.

Techniques: In Situ, Sterility, Derivative Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Adiponectin receptor T-cadherin emerges as a novel regulator of adipose stem cell quiescence and adipogenesis

doi: 10.3389/fcell.2025.1734183

Figure Lengend Snippet: Double immunofluorescent staining of human subcutaneous adipose tissue sections with antibodies against T-cadherin (green) and DPP4 (red); nuclei were counterstained with DAPI (blue). Images were acquired using a Zeiss LSM 780 confocal microscope and ZEN2010 software, shown at lower magnification (A) and higher magnification (B) . A thick arrow points to a group of cells expressing both T-cadherin and DPP4 in the interstitium; thin arrows mark cells expressing only T-cadherin; ovals encircle adipocytes. Scale bar 50 µm. (C) The table shows the percentage of T-cadherin–positive, DPP4 + cells and double-positive cells (DPP4 + /T-cadherin + ), quantified from adipose tissue sections of two healthy donors.

Article Snippet: Cells were detached from culture dishes using HyQTase Detachment Reagent (HyClone, GE Healthcare Life Sciences, United States) and stained with appropriate combinations of primary antibodies against: DPP4 (CD26 Antibody (MA2607), ThermoFisher Scientific, dilution 1:100), T-cadherin (ProSci, United States, #3583, dilution 1:100).

Techniques: Staining, Microscopy, Software, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Adiponectin receptor T-cadherin emerges as a novel regulator of adipose stem cell quiescence and adipogenesis

doi: 10.3389/fcell.2025.1734183

Figure Lengend Snippet: Light microscopy of MSCs (of the two to three passages) isolated from human subcutaneous adipose tissue of a healthy donor (A) and immunofluorescent staining with antibodies against T-cadherin (green) (B) . Arrows indicate cells with low or no T-cadherin expression, whereas cells exhibiting green fluorescence corresponding to T-cadherin are clearly visible. Scale bar, 50 µm. Light microscopy of human MSCs (C) and double immunofluorescent staining with antibodies against T-cadherin green, (E) and DPP4 red, (F) nuclei were counterstained with DAPI blue, (D) . Arrows in (C–F) indicate one and the same cell co-expressing T-cadherin and DPP4. Images were acquired using a Leica DMI 6000B microscope equipped with a Leica DFC7000T digital camera and LAS X software. Scale bar, 20 µm. (G) Representative flow cytometry plot showing T-cadherin and DPP4 distribution in cultured MSCs. The proportion of double-positive (DPP4 + /T-cadherin + ) cells was 30.4%; 6.15% expressed only T-cadherin, and 14% expressed only DPP4.

Article Snippet: Cells were detached from culture dishes using HyQTase Detachment Reagent (HyClone, GE Healthcare Life Sciences, United States) and stained with appropriate combinations of primary antibodies against: DPP4 (CD26 Antibody (MA2607), ThermoFisher Scientific, dilution 1:100), T-cadherin (ProSci, United States, #3583, dilution 1:100).

Techniques: Light Microscopy, Isolation, Staining, Expressing, Fluorescence, Microscopy, Software, Flow Cytometry, Cell Culture

Journal: Frontiers in Cell and Developmental Biology

Article Title: Adiponectin receptor T-cadherin emerges as a novel regulator of adipose stem cell quiescence and adipogenesis

doi: 10.3389/fcell.2025.1734183

Figure Lengend Snippet: Individual UMAP plots showing the expression levels and distribution of CDH13 (encoding T-cadherin) in control MSCs (A) and MSCs after 4 days of adipogenic induction (B) . UMAP plots demonstrating DPP4 expression in control MSCs (C) and MSCs after 4 days of adipogenic induction (D) . (E) RT-qPCR analysis of MSCs cultured in control medium or under adipogenic induction conditions showing the dynamics of T-cadherin mRNA expression. T-cadherin/ CDH13 expression decreased by day 4 in adipogenic medium and remained low through day 10. RT-qPCR data are shown as the mean ± SD. T-test. **р< 0.01 *p < 0.05 vs. control media in corresponding experimental day. Results are representative of three biologically independent experiments.

Article Snippet: Cells were detached from culture dishes using HyQTase Detachment Reagent (HyClone, GE Healthcare Life Sciences, United States) and stained with appropriate combinations of primary antibodies against: DPP4 (CD26 Antibody (MA2607), ThermoFisher Scientific, dilution 1:100), T-cadherin (ProSci, United States, #3583, dilution 1:100).

Techniques: Expressing, Control, Quantitative RT-PCR, Cell Culture

Journal: Frontiers in Cell and Developmental Biology

Article Title: Adiponectin receptor T-cadherin emerges as a novel regulator of adipose stem cell quiescence and adipogenesis

doi: 10.3389/fcell.2025.1734183

Figure Lengend Snippet: Integrated object. (A) FeaturePlot–UMAP-plot showing principal distribution of CDH13 gene expression (encoding for T-cadherin) in the integrated object; CDH13 expressing cells corresponds to Cluster 3 (more than 1-fold change of the average expression level); (B) FeaturePlot–UMAP-plot showing principal distribution of DPP4 gene expression (encoding for T-cadherin) in the integrated object; DPP4 expressing cells correspond to Cluster 3 (more than 1-fold change of the average expression level) (C) DimPlot–Integrated object UMAP-clustering. Sample proportion diagrams depict the ratio between the cell counts in the control MSC sample (Salmon) and in the MSC sample (Iris blue) after a 4-day induction of adipogenic differentiation within the Clusters. (D) DimPlot–Integrated object grouped by samples. CDH13 expression in the control MSC sample (Salmon) and MSC sample (Iris blue) after a 4-day induction of adipogenic differentiation. Cluster 3 predominantly contains cells from the control sample.

Article Snippet: Cells were detached from culture dishes using HyQTase Detachment Reagent (HyClone, GE Healthcare Life Sciences, United States) and stained with appropriate combinations of primary antibodies against: DPP4 (CD26 Antibody (MA2607), ThermoFisher Scientific, dilution 1:100), T-cadherin (ProSci, United States, #3583, dilution 1:100).

Techniques: Gene Expression, Expressing, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Adiponectin receptor T-cadherin emerges as a novel regulator of adipose stem cell quiescence and adipogenesis

doi: 10.3389/fcell.2025.1734183

Figure Lengend Snippet: Integrated object. FeaturePlot. Each cluster is denoted by color. Cluster 0 (Salmon) primarily contains cells expressing fibroblast markers and genes responsible for cell cycle regulation. Cluster 1 (Khaki) encompasses cells expressing preadipocyte-specific genes, such as CEBPB , PPARγ, CD36 and markers of mature adipocytes ( ADIPOQ , Perilipin1 , Perilipin4 ). In Cluster 2 (green), cells predominantly express genes related to mitosis. Cluster 3 (Blue) contains cells of interest with high level of T-cadherin expression, as well as classical MSC markers ( CD90 , PDGFR ), Wnt signaling genes , and DPP4 . In a separate remote Cluster 4 (Magenta), besides CDH13 , cells express Nestin , a marker of neural crest cells, and CD36 , a marker of adipocyte progenitors.

Article Snippet: Cells were detached from culture dishes using HyQTase Detachment Reagent (HyClone, GE Healthcare Life Sciences, United States) and stained with appropriate combinations of primary antibodies against: DPP4 (CD26 Antibody (MA2607), ThermoFisher Scientific, dilution 1:100), T-cadherin (ProSci, United States, #3583, dilution 1:100).

Techniques: Expressing, Marker

Journal: Frontiers in Cell and Developmental Biology

Article Title: Adiponectin receptor T-cadherin emerges as a novel regulator of adipose stem cell quiescence and adipogenesis

doi: 10.3389/fcell.2025.1734183

Figure Lengend Snippet: (A) DimPlot– GSE182158 object UMAP-clustering; (B) 2 cluster manual cell type annotation, the red oval marks cluster 2; (C) FeaturePlot–UMAP-plot showing principal distribution of CDH13 gene expression in the GSE182158 object; (D) FeaturePlot–UMAP-plot showing principal distribution of DPP4 gene expression (encoding for T-cadherin) in the GSE182158 object.

Article Snippet: Cells were detached from culture dishes using HyQTase Detachment Reagent (HyClone, GE Healthcare Life Sciences, United States) and stained with appropriate combinations of primary antibodies against: DPP4 (CD26 Antibody (MA2607), ThermoFisher Scientific, dilution 1:100), T-cadherin (ProSci, United States, #3583, dilution 1:100).

Techniques: Gene Expression

Journal: Frontiers in Cell and Developmental Biology

Article Title: Adiponectin receptor T-cadherin emerges as a novel regulator of adipose stem cell quiescence and adipogenesis

doi: 10.3389/fcell.2025.1734183

Figure Lengend Snippet: T-cadherin overexpression in human MSCs using lentiviral constructs comprising GFP for cell identification and sorting. Overexpression of T-cadherin in human MSCs using lentiviral constructs carrying either GFP alone (control) or T-cadherin–GFP, allowing subsequent cell identification and sorting. (A) T-cadherin overexpression (T+virus) was confirmed using immunofluorescent staining with antibodies against T-cadherin (red) and compared to Control virus cells (Contr virus). Cell nuclei were counterstained with DAPI (blue). Scale bar 50 m. T-cadherin overexpression was verified applying RT-qPCR (B) and Western blot (C) . GAPDH was used as the loading control (C) . Results from two biologically independent RT-qPCR and eight Western blot experiments are shown.

Article Snippet: Cells were detached from culture dishes using HyQTase Detachment Reagent (HyClone, GE Healthcare Life Sciences, United States) and stained with appropriate combinations of primary antibodies against: DPP4 (CD26 Antibody (MA2607), ThermoFisher Scientific, dilution 1:100), T-cadherin (ProSci, United States, #3583, dilution 1:100).

Techniques: Over Expression, Construct, Control, Virus, Staining, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Adiponectin receptor T-cadherin emerges as a novel regulator of adipose stem cell quiescence and adipogenesis

doi: 10.3389/fcell.2025.1734183

Figure Lengend Snippet: Elevated DPP4 expression in MSCs after lentiviral transduction in T-cadherin-overexpressing cells was verified using RT-qPCR (A) and Western blot (B) . β-tubulin was used as the loading control for Western blot analysis. Representative results from one of two biologically independent RT-qPCR and eight Western blot experiments are shown. ANOVA with multiple comparisons, **p < 0.01.

Article Snippet: Cells were detached from culture dishes using HyQTase Detachment Reagent (HyClone, GE Healthcare Life Sciences, United States) and stained with appropriate combinations of primary antibodies against: DPP4 (CD26 Antibody (MA2607), ThermoFisher Scientific, dilution 1:100), T-cadherin (ProSci, United States, #3583, dilution 1:100).

Techniques: Expressing, Transduction, Quantitative RT-PCR, Western Blot, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Adiponectin receptor T-cadherin emerges as a novel regulator of adipose stem cell quiescence and adipogenesis

doi: 10.3389/fcell.2025.1734183

Figure Lengend Snippet: T-cadherin–overexpressing MSCs demonstrated reduced proliferative capacity compared to control cells (either transduced with a control virus or untransduced donor-matched MSCs).

Article Snippet: Cells were detached from culture dishes using HyQTase Detachment Reagent (HyClone, GE Healthcare Life Sciences, United States) and stained with appropriate combinations of primary antibodies against: DPP4 (CD26 Antibody (MA2607), ThermoFisher Scientific, dilution 1:100), T-cadherin (ProSci, United States, #3583, dilution 1:100).

Techniques: Control, Transduction, Virus

Journal: Frontiers in Cell and Developmental Biology

Article Title: Adiponectin receptor T-cadherin emerges as a novel regulator of adipose stem cell quiescence and adipogenesis

doi: 10.3389/fcell.2025.1734183

Figure Lengend Snippet: Adipogenic differentiation in MSC cultures consisting of untransduced cells, cells transduced with either lentivirus for T-cadherin overexpression or control virus, following 14 days of culture in either control or adipogenic media. (A–F) Bright field and phase-contrast micrographs of MSC cultures after Oil Red O staining. Distinct orange lipid droplets are visible in individual cells and cell clusters in untransduced MSCs, and control virus–transduced MSCs, whereas T-cadherin–overexpressing MSCs show no detectable Oil Red O staining. Representative images from one of two biologically independent experiments are shown. Scale bar 250 μm. (G) Quantification of differentiated cells following Nile Red staining using AI-based image analysis and deep learning algorithms from the NIS.ai module, showing the ratio of differentiated adipocytes to total cells. (H) Western blot assessment of an early adipocyte differentiation marker CEBP Beta in lysates of MSCs [WT—untransduced cells, cells transduced with either lentivirus for T-cadherin overexpression (T+) or control virus (GFP)] cultured in adipogenic or control condition for 9 days. For loading control, β-tubulin antibody was used. (d0 – start of the experiment; d6, d9—days following adipogenic induction). (I) Densitometric quantification of Western blot analysis of CEBP Beta levels.

Article Snippet: Cells were detached from culture dishes using HyQTase Detachment Reagent (HyClone, GE Healthcare Life Sciences, United States) and stained with appropriate combinations of primary antibodies against: DPP4 (CD26 Antibody (MA2607), ThermoFisher Scientific, dilution 1:100), T-cadherin (ProSci, United States, #3583, dilution 1:100).

Techniques: Transduction, Over Expression, Control, Virus, Staining, Western Blot, Marker, Cell Culture

Journal: Frontiers in Cell and Developmental Biology

Article Title: Adiponectin receptor T-cadherin emerges as a novel regulator of adipose stem cell quiescence and adipogenesis

doi: 10.3389/fcell.2025.1734183

Figure Lengend Snippet: Human MSC culture after lentivirus transduction for T-cadherin overexpression (representative images for the cell culture transduced with lentivirus for T-cadherin overexpression). Images of MSCs were captured through a long-term live-cell imaging over a period of 10 days. Transduced cells were identified by red fluorescence (mKate fluorescence with emission maximum 633 nm, pseudo-colored in yellow) (A) . Cells displaying mKate fluorescence and overexpressing T-cadherin are encircled in red, while the neighboring untransduced cells serve as internal control and are encircled in yellow (A,B) . Panels (C,D) show the same field of view as in (A,B) after 10 days [ (C) —phase contrast; (D) green fluorescence channel]. Nile Red staining was used for visualization of non-polar (neutral) lipids (green fluorescence channel) (D) . Images were acquired using a microscope NIS-Elements (Nikon) and ImageJ software (NIH, Bethesda, MD, United States). Scale bar 100 μm. (E) Quantification of cells undergoing adipogenic differentiation in MSC cultures transduced with lentivirus for T-cadherin overexpression or the control virus (T+vir culture and contr.vir culture). **p = 0.0008.

Article Snippet: Cells were detached from culture dishes using HyQTase Detachment Reagent (HyClone, GE Healthcare Life Sciences, United States) and stained with appropriate combinations of primary antibodies against: DPP4 (CD26 Antibody (MA2607), ThermoFisher Scientific, dilution 1:100), T-cadherin (ProSci, United States, #3583, dilution 1:100).

Techniques: Transduction, Over Expression, Cell Culture, Live Cell Imaging, Fluorescence, Control, Staining, Microscopy, Software, Virus