Journal: Neurochemistry international

Article Title: Increased spinal adenosine after subacute cervical injury correlates with sustained upregulation of CD39 and CD73 in microglia

doi: 10.1016/j.neuint.2025.106030

Figure Lengend Snippet: CD39 mRNA was assessed in controls (uninjured) or 1-, 2- and 8-weeks post-C2 hemisection in isolated microglia and microglia-free homogenates at (A) and below (B) injury. (C) Flow cytometry analysis of CD39 protein. Example of CD39 in C1–C3 microglia in uninjured (upper left); 1-week post-injury (upper right); histogram of fluorescence intensity (bottom). Negative controls are cells stained with CD11b-PE antibody and secondary-Alexa405 antibody without CD39 antibody. (D) CD39 protein analysis for experimental groups. MFI, median fluorescent intensity. Controls (uninjured rats) were included for each time points 1-, 2-, 8-weeks after injury. Because there were no differences in control rats, data were pooled together into one ctrl bar. N = 4–8/group; **p < 0.01 vs ctrl, *p < 0.05 vs ctrl; +++p < 0.001 vs microglia.

Article Snippet: 2 , CD11b-PE (Miltenyi) , CD39 (Proteintech) , Goat anti-rabbit-Alexa 405 (Invitrogen).

Techniques: Isolation, Flow Cytometry, Fluorescence, Staining, Control

Journal: Neurochemistry international

Article Title: Increased spinal adenosine after subacute cervical injury correlates with sustained upregulation of CD39 and CD73 in microglia

doi: 10.1016/j.neuint.2025.106030

Figure Lengend Snippet: CD73 mRNA was assessed in controls (uninjured) or 1-, 2- and 8-weeks after C2 hemisection in isolated microglia and microglia-free homogenates at (A) and below (B) injury. (C) Flow cytometry analysis of CD73 protein. Example of CD73 in C1–C3 microglia in controls (top left) and 1-week post-injury (top right); with histogram of fluorescence intensity (bottom). Negative controls are cells stained with CD11b-PE antibody and secondary-Alexa405 antibody without CD73 antibody. (D) CD39 protein analysis for each experimental group. MFI, median fluorescent intensity. Controls (uninjured rats) were included for each time points 1-, 2-, 8-weeks after injury. Because there were no differences in control rats, data were pooled together into one ctrl bar N = 4–8/group C. CD39 mRNA expression in microglia in control and injured animals. N = 4/group; **p < 0.01 vs ctrl, *p < 0.05 vs ctrl; +++p < 0.001 vs microglia.

Article Snippet: 2 , CD11b-PE (Miltenyi) , CD39 (Proteintech) , Goat anti-rabbit-Alexa 405 (Invitrogen).

Techniques: Isolation, Flow Cytometry, Fluorescence, Staining, Control, Expressing

Journal: Neurochemistry international

Article Title: Increased spinal adenosine after subacute cervical injury correlates with sustained upregulation of CD39 and CD73 in microglia

doi: 10.1016/j.neuint.2025.106030

Figure Lengend Snippet: CD39 mRNA levels were correlated with the expression of other microglial genes after SCI at (A) and below (B) the injury site. Similar correlation analysis was performed for CD73 at (C) and below (D) injury. The strongest correlation was observed with P2Y12 in both regions.

Article Snippet: 2 , CD11b-PE (Miltenyi) , CD39 (Proteintech) , Goat anti-rabbit-Alexa 405 (Invitrogen).

Techniques: Expressing

Journal: Neurochemistry international

Article Title: Increased spinal adenosine after subacute cervical injury correlates with sustained upregulation of CD39 and CD73 in microglia

doi: 10.1016/j.neuint.2025.106030

Figure Lengend Snippet: (A) Adenosine was measured in ventral C3–C6 spinal segments in intact animals (ctrl) and 2- and 9-weeks post C2 hemisection (SCI). N = 6–8/group. **p < 0.01 vs ctrl. (B) There was no correlation between spinal adenosine and non-microglial CD39 and CD73 expression. However, adenosine levels strongly correlated with microglial CD39 and CD73 both on the protein (C) and mRNA levels (D) . Because spinal adenosine and CD73/CD39 expression were analyzed in different sets of animals we used mean (SE) for correlation analyses.

Article Snippet: 2 , CD11b-PE (Miltenyi) , CD39 (Proteintech) , Goat anti-rabbit-Alexa 405 (Invitrogen).

Techniques: Expressing

Journal: Neurochemistry international

Article Title: Increased spinal adenosine after subacute cervical injury correlates with sustained upregulation of CD39 and CD73 in microglia

doi: 10.1016/j.neuint.2025.106030

Figure Lengend Snippet: SCI-associated cell damage, tissue hypoxia, and inflammation increase extracellular ATP promoting inflammation and neuronal damage. Microglia upregulation of CD39 (converting ATP to ADP to AMP) and CD73 (converting AMP to adenosine (ADO)) increases adenosine levels determining the magnitude and mechanism of mAIH-induced phrenic motor plasticity. attenuating microglial inflammatory responses. Conversely, increasing levels of adenosine shift the serotonin/adenosine balance. Figure was created in Biorender .

Article Snippet: 2 , CD11b-PE (Miltenyi) , CD39 (Proteintech) , Goat anti-rabbit-Alexa 405 (Invitrogen).

Techniques:

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: Cancer Research

Article Title: Selective Depletion of CD4+CD25+Foxp3+ Regulatory T Cells by Low-Dose Cyclophosphamide Is Explained by Reduced Intracellular ATP Levels

doi: 10.1158/0008-5472.can-10-0283

Figure Lengend Snippet: Figure 2. miR-142-3p and CD39 contributes to the low level of ATP in Treg cells. A, Treg cells and conventional T cells were isolated from mice and humans. The expression of miR-142-3p was analyzed by Northern blot. B, the level of miR-142-3p influences the concentration of ATP in Treg cells. The isolated murine or human Treg cells were transfected with miR-142-3p (left) or miR-142-3p inhibitor (right). Twenty-four hours later, the intracellular ATP levels were measured with the somatic cell ATP assay kit. C, the activity of CD39 influences the concentration of ATP in Treg cells. The isolated murine or human Treg cells were treated with CD39 inhibitor ARL67156 (left) or recombinant CD39 (right). Twenty-four hours later, the intracellular ATP levels were measured with the somatic cell ATP assay kit.

Article Snippet: In parallel, we also cultured CD4+CD25− T cells isolated from mouse spleen or human PBMCs in the presence or absence of recombinant mouse or human CD39 (0.5 μg/mL; R & D Systems).

Techniques: Isolation, Expressing, Northern Blot, Concentration Assay, Transfection, ATP Assay, Activity Assay, Recombinant

Journal: Cardiovascular Diabetology

Article Title: Elevated glucose levels increase vascular calcification risk by disrupting extracellular pyrophosphate metabolism

doi: 10.1186/s12933-024-02502-w

Figure Lengend Snippet: Impact of high glucose on extracellular pyrophosphate metabolism. Aortic smooth muscle cells were cultured for one month in media containing either low (1 g/L) or high (4.5 g/L) glucose. A Measurement of extracellular pyrophosphate levels. B Extracellular pyrophosphate-to-ATP ratio. C , D Analysis of the gene expression of key enzymes involved in extracellular pyrophosphate metabolism, including eNTPD1, eNPP1, and TNAP, from isolated total RNA. (E) Immunoblot analysis of proteins associated with extracellular pyrophosphate metabolism. F , G Quantification of protein levels via ELISA, highlighting significant differences. The data are shown as the mean ± SEM, with data derived from 4 independent experiments, each containing 4 replicate plates. Statistical significance was determined via Student’s t test, with asterisks denoting significance levels: * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: The recombinant enzymes eNPP1 (catalog number 6136-EN) and eNTPD1 (catalog number 4397-EN) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Cell Culture, Gene Expression, Isolation, Western Blot, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: Cardiovascular Diabetology

Article Title: Elevated glucose levels increase vascular calcification risk by disrupting extracellular pyrophosphate metabolism

doi: 10.1186/s12933-024-02502-w

Figure Lengend Snippet: High glucose levels impair the pyrophosphate-to-phosphate ratio. Aortic smooth muscle cells were incubated for one month in medium containing 1 g/L or 4.5 g/L glucose. A Autoradiograph displaying representative products from the hydrolysis of ATP (1 µmol/L ATP, 10 µCi/mL [γ 32 Pi]ATP) incubated with or without recombinant eNPP1 (ectonucleotide pyrophosphatase/phosphodiesterase 1) or eNTPD1 (ectonucleoside triphosphate diphosphohydrolase 1) enzymes. Enzymatic hydrolysis generated radiolabeled 32 PPi (32-pyrophosphate) and 32 Pi (32-phosphate), which, alongside unreacted [γ 32 Pi]ATP, were separated by thin-layer chromatography (TLC), as detailed in the section. B Representative time course of ATP hydrolysis showing the products released over time. C Synthesis of the pyrophosphate 32 PPi via hydrolysis of [γ 32 Pi]ATP (10 µCi/mL; 1 µmol/L ATP) in the absence or presence of 100 µmol/L SBI245 (a specific TNAP inhibitor) or inorganic pyrophosphatase (PPase). D The pyrophosphate-to-phosphate ( 32 PPi/ 32 Pi) ratio was quantified following hydrolysis of [γ 32 Pi]ATP (10 µCi/mL; 1 µmol/L ATP) under various conditions: in the absence of inhibitors (Control), in the presence of an ectonucleoside triphosphate diphosphohydrolase (eNTPD) inhibitor (INH, 200 µmol/L), or with the recombinant enzymes eNPP1 and eNTPD1 (100 ng/mL). Experiments were conducted in media containing either physiological (1 g/L) or elevated (4.5 g/L) glucose concentrations. D) Synthesis of 32 PPi by hydrolysis of [γ 32 Pi]ATP (10 µCi/mL and 1 µmol/L ATP). E Hydrolysis of 32 PPi (10 µCi/mL and 5 µmol/L PPi). The results are shown as the mean ± SEM (4 independent experiments with 4 plates per experiment). Student’s t test ( E, F ) or one-way ANOVA with Tukey’s post hoc test ( C, D ) was used for statistical analysis. Asterisks indicate a statistically significant difference compared with the control group: * P < 0.05; *** P < 0.001. ### Indicates a value of P < 0.001 compared with the control group (1 g/L)

Article Snippet: The recombinant enzymes eNPP1 (catalog number 6136-EN) and eNTPD1 (catalog number 4397-EN) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Incubation, Autoradiography, Recombinant, Generated, Thin Layer Chromatography, Control

Journal: Cardiovascular Diabetology

Article Title: Elevated glucose levels increase vascular calcification risk by disrupting extracellular pyrophosphate metabolism

doi: 10.1186/s12933-024-02502-w

Figure Lengend Snippet: STZ-treated rats exhibit impaired extracellular pyrophosphate metabolism in the aortic wall. A A representative time course of ATP hydrolysis was conducted using a 1 µmol/L ATP solution containing 10 µCi/mL [γ- 32 P]ATP as a radiotracer. The products of hydrolysis, 32 PPi (32-pyrophosphate), 32 Pi (32-phosphate), and [γ- 32 P]ATP-, were separated and quantified via thin layer chromatography, as outlined in the section. B The synthesis of pyrophosphate (PPi) was analyzed by hydrolyzing 1 µmol/L ATP containing 10 µCi/mL [γ- 32 P]ATP as a radiotracer. The reactions were carried out in the absence or presence of either a specific TNAP inhibitor (SBI-425) or inorganic pyrophosphatase (PPase). C The ratio of 32 PPi to 32 Pi generated by ATP hydrolysis was calculated to assess the efficiency and specificity of pyrophosphate synthesis. D The synthesis of 32 PPi was evaluated by hydrolyzing 1 µmol/L ATP containing 10 µCi/mL [γ- 32 P]ATP. E The release of 32 Pi was measured following the hydrolysis of 5 µmol/L pyrophosphate, which contained 10 µCi/mL 32 PPi as a radiotracer. F Quantification of protein levels via ELISA. G , H Total RNA was isolated from rat aortas to evaluate the expression levels of key enzymes involved in extracellular pyrophosphate metabolism, including eNTPD1 (ectonucleoside triphosphate diphosphohydrolase 1), eNPP1 (ectonucleotide pyrophosphatase/phosphodiesterase 1), and tissue-nonspecific alkaline phosphatase (TNAP) (panel G . Additionally, the expression of calcification-related proteins, such as matrix Gla protein (MGP) and osteopontin (OPN), was assessed (panel H). The data are shown as the mean ± SEM and represent data from 12–16 independent aortas. Statistical analyses were performed via Student’s t test. Asterisks indicate a significant difference with *** P < 0.001

Article Snippet: The recombinant enzymes eNPP1 (catalog number 6136-EN) and eNTPD1 (catalog number 4397-EN) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Thin Layer Chromatography, Generated, Enzyme-linked Immunosorbent Assay, Isolation, Expressing

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