β methylene diphosphate sodium salt apcp  (Tocris)

Journal: Cell Communication and Signaling : CCS

Article Title: CD73/adenosine axis exerts cardioprotection against hypobaric hypoxia-induced metabolic shift and myocarditis in a sex-dependent manner

doi: 10.1186/s12964-024-01535-8

Figure Lengend Snippet: CD73/adenosine axis mediates immunomodulation during HH in a sex-dependent manner. A - B ELISA analysis of Extracellular Adenosine level and CD73 activity ( n = 4 mice/group). C Relative mRNA levels of adenosine receptors (A1, A2a, A2b, A3 and n = 4mice/group). D-F Representative immunohistochemical staining and graphical presentation of macrophage CD86 and CD206-positive cells assessed from the myocardial sections ( n = 4–6 hearts per group) Scale bar, 50 μm. G-K Graphical presentation of sera cardiac troponin I (cTnI) concentrations and inflammatory cytokines; Interleukin (IL)-1β, IL-18, IL-10, and transforming growth factor (TGF)-β concentrations assessed by ELISA and RT-qPCR using myocardia lysates. All ELISA were performed in triplicates ( n = 5–6 mice per group). M = male; MH = male + HH; F = female; FH = female + HH; MI = male + APCP; FI = female + APCP; MHI = male + HH + APCP; FHI = female + HH + APCP. Data are presented as mean ± SEM; *** p < 0.001; ** p < 0.01; * p < 0.05

Article Snippet: CD73 inhibitor Adenosine 5′-(α, β-methylene) diphosphate sodium salt (APCP) was purchased from Tocris Bio-Techne, and the prostatic acid phosphatase (PAP) inhibitor – Benzylphosphonic acid (GB57232) (BPA) was purchased from GlpBio Technology Inc. All the drugs were prepared according to the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Activity Assay, Immunohistochemical staining, Staining, Quantitative RT-PCR

Journal: Cell Communication and Signaling : CCS

Article Title: CD73/adenosine axis exerts cardioprotection against hypobaric hypoxia-induced metabolic shift and myocarditis in a sex-dependent manner

doi: 10.1186/s12964-024-01535-8

Figure Lengend Snippet: Hyperactivity of CD73/adenosine axis promotes myocardial metabolic shift in a sex-dependent manner during HH and protein expression of the other ectonucleotidases. A - B Representative Immunofluorescence and graphical presentation of the co-staining of F4/80 and Glut1 with nuclei (DAPI). Color channels were adjusted in the merged images to enhance the visualization of all the respective fluorescence dyes. Scale bar, 50 μm. C - E Representative Oil Red O (ORO) and Periodic Acid Schiff (PAS) staining of myocardial sections and their respective graphical presentations showing lipid and glycogen deposition percentages ( n = 4–6 sections per 4–6 mice per group) Scale bar, 50 μm. F - H Representative Immunoblotting of CD36 and Glut1 and their respective Graphical plots; each blot band in the representative blot is an independent biological sample ( n = 3 hearts per group). I - L Representative Immunoblotting of CD73, TNAP, and PAP and their respective Graphical plots; each blot band in the representative blot is an independent biological sample ( n = 3 hearts per group). M = male; MH = male + HH; F = female; FH = female + HH; MI = male + APCP; FI = female + APCP; MHI = male + HH + APCP; FHI = female + HH + APCP. Data are expressed as mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: CD73 inhibitor Adenosine 5′-(α, β-methylene) diphosphate sodium salt (APCP) was purchased from Tocris Bio-Techne, and the prostatic acid phosphatase (PAP) inhibitor – Benzylphosphonic acid (GB57232) (BPA) was purchased from GlpBio Technology Inc. All the drugs were prepared according to the manufacturer's instructions.

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: CD73/adenosine axis exerts cardioprotection against hypobaric hypoxia-induced metabolic shift and myocarditis in a sex-dependent manner

doi: 10.1186/s12964-024-01535-8

Figure Lengend Snippet: Double inhibition of PAP and CD73 in male mice, mitigated glycolytic shift and proinflammatory response. A - C Representative Immunoblotting of CD73, TNAP, and PAP and their respective Graphical plots; each blot band in the representative blot is an independent biological sample ( n = 3 hearts per group). E ELISA analysis of Extracellular Adenosine level (n = 4 mice/group). F - H Representative Oil Red O (ORO) and Periodic Acid Schiff (PAS) staining of myocardial sections and their respective graphical presentations showing lipid and glycogen deposition percentages ( n = 4–6 sections per 4–5 mice per group) Scale bar, 50 μm. I - K Representative Immunoblotting of CD36 and Glut1 and their respective Graphical plots; each blot band in the representative blot is an independent biological sample ( n = 3 hearts per group). L - N Representative immunohistochemical staining and graphical presentation of macrophage CD86 and CD206-positive cells assessed from the myocardial sections ( n = 4–5 hearts per group) Scale bar, 50 μm. O - Q Inflammatory cytokines; Interleukin TNF-α, IL-10, and transforming growth factor (TGF)-β concentrations assessed by ELISA using myocardia lysates. All ELISA were performed in triplicates ( n = 4–5 mice per group). M = male; MH = male + HH; F = female; FH = female + HH; MI = male + APCP; FI = female + APCP; MHI = male + HH + APCP; FHI = female + HH + APCP. Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: CD73 inhibitor Adenosine 5′-(α, β-methylene) diphosphate sodium salt (APCP) was purchased from Tocris Bio-Techne, and the prostatic acid phosphatase (PAP) inhibitor – Benzylphosphonic acid (GB57232) (BPA) was purchased from GlpBio Technology Inc. All the drugs were prepared according to the manufacturer's instructions.

Techniques: Inhibition, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemical staining

Journal: Journal of leukocyte biology

Article Title: Adenosine from a biologic source regulates neutrophil extracellular traps (NETs)

doi: 10.1002/JLB.3VMA0918-374R

Figure Lengend Snippet: MSCs were pretreated overnight with 100 μM APCP, a nonhydrolizable inhibitor of CD73. HPLC analysis of conditioned media showed no ADO production without supplementation of the CD73 substrate AMP (A). MSC treatment with APCP had almost complete impairment of adenosine production from extracellular AMP (A). Quantification of SYTOX green fluorescence indicates that APCP impairs the ability of MSCs to inhibit NET production (B; n = 10). The relative inhibition percentage of APCP–MSC-treated PMN group was significantly reduced compared to the CD73 competent MSC-treated group by human neutrophil elastase-DNA ELISA (C; n = 7). Data are expressed as mean ± SD, *P < 0.05, ***P < 0.001. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc analysis

Article Snippet: Some MSCs were treated for 12 hours with and without 100 μ M of the irreversible CD73 inhibitor, adenosine 5 ′ ( α , β -methylene) diphosphate sodium salt (APCP; #363310; Tocris, Minneapolis, MN).

Techniques: Fluorescence, Inhibition, Enzyme-linked Immunosorbent Assay