Journal: Journal of autoimmunity

Article Title: P2RX7 gene variants associate with altered inflammasome assembly and reduced pyroptosis in chronic nonbacterial osteomyelitis (CNO).

doi: 10.1016/j.jaut.2024.103183

Figure Lengend Snippet: Fig. 2. P2X7 determines pyroptosis and cytokine release. Pyroptosis, IL-1β and IL-18 release in response to priming (LPS) and stimulation (LPS + BzATP) were investigated in P2X7-deficient cells (mono- 3F2 and bi-allelic 4G8 deletion; A-C) and in THP-1 derived 4G8 macrophages reconstituted with wild-type P2X7 expression vectors (D–F). A-C) Kruskal Wallis test and Dunn’s multiple comparison (result shown in black print; A: N ≥6, B/C: N ≥8). A-C) Wilcoxon tests comparing THP1 to 3F2 cells and THP1 to 4G8 cells (result shown in blue print). D-F) Wilcoxon tests (D: N = 6, E/F: N ≥8); *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.001.

Article Snippet: THP-1 cells were treated overnight with PMA (2.5 ng/ml, Sigma- A. Charras et al. Journal of Autoimmunity 144 (2024) 103183 Aldrich) to generate macrophage-like cells, followed by priming (1 μg/ ml LPS, Sigma-Aldrich) for 3h and treated with LPS, LPS/ATP (1 mM, Invivogen), LPS/BzATP (150/300 μM, Alomone), or LPS/nigericin (5/ 10 μM) as indicated.

Techniques: Derivative Assay, Expressing, Comparison

Journal: Neurobiology of disease

Article Title: P2RX7 modulates the function of human microglia-like cells and mediates the association of IL18 with Alzheimer’s disease traits

doi: 10.1016/j.nbd.2025.107106

Figure Lengend Snippet: Extracellular ATP induces IL1β and IL18 gene expression upregulation in a P2RX7-dependent manner. A) IL1β , IL18 , and NLRP3 expression levels were measured with qPCR from MDMi treated with DMSO alone (Ctrl), 1 mM ATP (ATP), or 1 mM ATP plus 10 μM A740003 (ATP + A74). Statistics were determined with a repeated measure one-way paired ANOVA with Newman-Keuls multiple comparisons test. N = 14. B) IL-1β ( N = 6) and IL-18 ( N = 5) protein detection in supernatants of MDMi treated with DMSO alone (Ctrl), 1 mM ATP (ATP), or 1 mM ATP plus 10 μM A740003 (ATP + A74). Statistics were determined with a two-way paired ANOVA. C) IL1β , IL18 , and NLRP3 expression levels were measured with qPCR from MDMi treated with DMSO alone (Ctrl), or 500 μM bzATP (bzATP). Statistics were determined with a paired t -test. N = 9. ns = p > 0.05, * p ≤ 0.05, ** ≤ 0.01,*** ≤ 0.001, **** ≤ 0.0001.

Article Snippet: ATP was purchased from Theromfisher (#R0441) and bzATP was purchased from Tocris Bioscience (#3312).

Techniques: Gene Expression, Expressing

Journal: Neurobiology of disease

Article Title: P2RX7 modulates the function of human microglia-like cells and mediates the association of IL18 with Alzheimer’s disease traits

doi: 10.1016/j.nbd.2025.107106

Figure Lengend Snippet: ATP-mediated P2RX7 activation reduces MDMi Aβ1–42 uptake abilities. A) Mean fluorescence intensity (MFI) of HyLite Fluor 647-labeled Aβ1–42 peptide uptake normalized to DAPI MFI in MDMi treated with DMSO alone (Ctrl), 1 mM ATP (ATP), or 1 mM ATP and 10 μM A740003 (ATP + A74), measured by fluorescence on a microplate reader. Data are represented relative to the control condition. Statistics were determined with a repeated measure one-way paired ANOVA with Newman-Keuls multiple comparisons test. N = 26. B) MFI of HyLite Fluor 647-labeled Aβ1–42 peptide uptake normalized to DAPI MFI in MDMi treated with DMSO alone (Ctrl), or 500 μM bzATP (bzATP), as measured by fluorescence on a microplate reader. Data are represented relative to the control condition. Statistics were determined with a paired t-test. N = 14. C) Mean fluorescence intensity (MFI) of HyLite Fluor 647-labeled Aβ1–42 per DAPI+ cell in MDMi imaged using confocal microscopy. MDMi were treated with DMSO alone (Ctrl), 1 mM ATP (ATP), or 1 mM ATP and 10 μM A740003 (ATP + A74). Statistics were determined with a repeated measure one-way paired ANOVA with Newman-Keuls multiple comparisons test. N = 12. D) Representative images of MDMi treated with DMSO alone (Ctrl), 1 mM ATP (ATP), or 1 mM ATP and 10 μM A740003 (ATP + A74) and stained for P2RX7 (green) and DAPI (blue) and used to measure uptake of HyLite Fluor 647-labeled Aβ1–42 peptide (red). Images were processed in CellProfiler. ns = p > 0.05, *p ≤ 0.05, ** ≤ 0.01,*** ≤ 0.001, **** ≤ 0.0001.

Article Snippet: ATP was purchased from Theromfisher (#R0441) and bzATP was purchased from Tocris Bioscience (#3312).

Techniques: Activation Assay, Fluorescence, Labeling, Control, Confocal Microscopy, Staining

Journal: Journal of Neurochemistry

Article Title: Continued P2X7 activation leads to mitochondrial fission and compromising microglial phagocytosis after subarachnoid haemorrhage

doi: 10.1111/jnc.15712

Figure Lengend Snippet: Continued BzATP stimulation resulted in a gradual decrease in primary microglial phagocytosis. Microglia were continuously stimulated with BzATP for different periods (0, 1, 2, 6, 12, 24, 36, and 48 h), and pHrodo E. coli particles was incubated for 45 min. (a) the pHrodo intensity (green) and CCK8 (orange) absorbance was scanned with a fluorometer; (b) representative pictures under a fluorescence microscope (blue, DAPI). Scale bar = 100 μm. N denotes the number of independent cell culture preparations, n = 5–6 per group. Brown‐Forsythe ANOVA and Dunnett's T3 test, # represents the comparison from ctrl. Microglia were stimulated with BzATP for different periods (0, 2, 6, 24 h), and then (c) the pHrodo E. coli particles or (d) DiO‐labelled cell debris was added and incubated for 45 min. Then, the number and fluorescence intensity of positive cells were evaluated by flow cytometry, and the phagocytic index was calculated. N denotes the number of independent cell culture preparations, n = 5–6 per group. Microglia were pretreated with the P2X7‐specific inhibitor JNJ‐47965567 (JNJ for short in the panel) and were then stimulated with BzATP for 24 h, and (e) the pHrodo E. coli particles and (f) DiO‐labelled cell debris were incubated for 45 min before the phagocytic index was observed with flow cytometry. N denotes the number of independent cell culture preparations, n = 4–6 per group. Brown–Forsythe ANOVA and Dunnett's T3 test, # represents the comparison between 24 h and Ctrl. NC, negative control; Exact p values noted; ns, not significant; ## p < 0.01; ### p < 0.001; * p < 0.05; ** p < 0. 01; *** p < 0.001; **** p < 0.0001.

Article Snippet: More than one study found that 100 μM BzATP (Topscience # TP2226, Shanghai) can significantly activate P2X7 and change the cell membrane potential, so we also used the same concentration (Coddou et al., ).

Techniques: Incubation, Fluorescence, Microscopy, Cell Culture, Comparison, Flow Cytometry, Negative Control, FACS

Journal: Journal of Neurochemistry

Article Title: Continued P2X7 activation leads to mitochondrial fission and compromising microglial phagocytosis after subarachnoid haemorrhage

doi: 10.1111/jnc.15712

Figure Lengend Snippet: Excessive microglial mitochondrial fission after P2X7 activation may be associated with decreased phagocytosis. (a) Microglia were pretreated with JNJ‐47965567 or Mdivi‐1 followed by BzATP stimulation for 24 h and then labelled with MitoTracker red. (b) Branch length mean and (c) summed branches length mean were counted with MiNA plugin. N denotes the number of pictures, n = 109–208 of each group. (d) Display of all mitochondrial branch volumes in each group. Each dot indicates one mitochondrion. (e) The number of branches was analysed after 3D reconstruction. N denotes the number of microglia, n = 8–17 per group. (f, g & j, k) microglial phagocytosis after 24 h of BzATP stimulation was observed by flow cytometry. Microglia were pretreated with Mdivi‐1 to prevent mitochondrial fission, and the phagocytic index was calculated. N denotes the number of independent cell culture preparations, n = 4–6 per group. (h, i) Western blotting for shRNA interference efficiency, comparisons between WT and shRNA. N denotes the number of pictures, n = 4 of each group. Kruskal–Wallis test with Dunn's test in panels (b–d) but Brown–Forsythe ANOVA and Dunnett's T3 test in panel (e–k). JNJ‐47965567, JNJ for short in the panel; # represents the comparison between BzATP+Mdivi‐1 and BzATP; exact p values noted; ns, not significant; # p < 0.05; ## p < 0.01; #### p < 0.0001; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001

Article Snippet: More than one study found that 100 μM BzATP (Topscience # TP2226, Shanghai) can significantly activate P2X7 and change the cell membrane potential, so we also used the same concentration (Coddou et al., ).

Techniques: Activation Assay, Flow Cytometry, Cell Culture, Western Blot, shRNA, Comparison, FACS, Knockdown, Fluorescence Microscopy

Journal: Journal of Neurochemistry

Article Title: Continued P2X7 activation leads to mitochondrial fission and compromising microglial phagocytosis after subarachnoid haemorrhage

doi: 10.1111/jnc.15712

Figure Lengend Snippet: Excessive mitochondrial fission leads to decreased membrane potential (MtMP) and impaired ATP production. Microglia were treated with BzATP for 24 h with or without the presence of JNJ‐47965567 or Mivi‐1. Mitochondrial membrane potential was assessed with TMRE & JC‐10 staining. (a, b) Primary microglial MtMP analysed by TMRE staining. FCCP was used as positive control. (c, d) BV‐2 microglial MtMP analysed by TMRE staining. (e) The ratio of PE/FITC in JC‐10 staining was calculated. (f) ATP production in microglia in the presence of P2X7‐activated mitochondrial fission, and Mdivi‐1 was used as a control. (g, h) Primary microglial ROS production analysed by DCFH. (j, k) BV‐2 microglial ROS production analysed by DCFH. ROSUP was used as a positive control. N denotes the number of independent cell culture preparations, n = 4–6 per group. Brown–Forsythe ANOVA and Dunnett's T3 test. JNJ‐47965567, JNJ for short in the panel; exact p values noted; * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: More than one study found that 100 μM BzATP (Topscience # TP2226, Shanghai) can significantly activate P2X7 and change the cell membrane potential, so we also used the same concentration (Coddou et al., ).

Techniques: Membrane, Staining, Positive Control, Control, Cell Culture, shRNA, FACS

Journal: Journal of Neurochemistry

Article Title: Continued P2X7 activation leads to mitochondrial fission and compromising microglial phagocytosis after subarachnoid haemorrhage

doi: 10.1111/jnc.15712

Figure Lengend Snippet: Activation of microglial P2X7 following BzATP stimulation triggers Ca 2+ influx and activates CaN activity. The microglia were pre‐loaded with Fluo‐8 AM, a Ca 2+ binding dye and stimulated with BzATP, the fluorescence intensity was real‐time monitored by a fluorometer for 30 min. (a) Fluo‐8 fluorescence intensity readings before and after BzATP stimulation. N denotes the number of independent cell culture preparations, n = 4 per group. Two‐way ANOVA with Dunnett's test. (b) Representative Fluo‐8 fluorescence pictures. Scale bar = 50 μm. (c, d) Western blotting was used to detect the changes in CaN protein expression in microglia stimulated with BzATP, and β‐Actin was used as an internal reference. N denotes the number of independent cell culture preparations, n = 4 per group. One‐way ANOVA with Tukey's test. (e) CaN activity was detected at different time points (0, 2, 6, and 24 h) after the primary microglia were stimulated with BzATP. N denotes the number of independent cell culture preparations, n = 6 per group. (f) CaN activity changes were observed under specific inhibition of P2X7 at 24 h BzATP exposure, and FK506 was used as a negative control. N denotes the number of independent cell culture preparations, n = 6 per group. Brown–Forsythe ANOVA with Dunnett's T3 test in panel (e, f). JNJ‐47965567, JNJ for short in the panel; RFU, relative fluorescence units; exact p values noted; ns, not significant; ## p < 0.001; ### p < 0.001; * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: More than one study found that 100 μM BzATP (Topscience # TP2226, Shanghai) can significantly activate P2X7 and change the cell membrane potential, so we also used the same concentration (Coddou et al., ).

Techniques: Activation Assay, Activity Assay, Binding Assay, Fluorescence, Cell Culture, Western Blot, Expressing, Inhibition, Negative Control

Journal: Journal of Neurochemistry

Article Title: Continued P2X7 activation leads to mitochondrial fission and compromising microglial phagocytosis after subarachnoid haemorrhage

doi: 10.1111/jnc.15712

Figure Lengend Snippet: P2X7 activation promotes DRP1 S637 dephosphorylation by CaN. Microglia was treated with BzATP for 24 h in the presence of FK506 or not, then labelled with MitoTracker red. (a) Western blotting was used to assess DRP1 total protein, S616 phosphorylation and S637 phosphorylation. (b) Relative intensity analysis of S637 phosphorylation of DRP1. N denotes the number of independent cell culture preparations, n = 5 per group, Brown–Forsythe ANOVA with Dunnett's T3 test. (c) Representative pictures of mitochondria from two groups. (d) Branch length mean and summed branches length mean were counted with MiNA plugin. N denotes the number of pictures, n = 98 and 79, respectively, two‐sided Mann–Whitney U test. (e, f) Microglia was stimulated with BzATP for 24 h after CaN was inhibited by FK506. Microglial phagocytosis was assessed by flow cytometry after CaN was inhibited. (g) Western blotting for S637D mutation efficiency. (h, i) Microglial phagocytosis was assessed by flow cytometry after S637D mutation. N denotes the number of independent cell culture preparations, n = 3–5 per group, one‐way ANOVA with Tukey's test. NC, negative control; JNJ‐47965567, JNJ for short in the panel; Exact p values noted; ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001, **** p < 0.05.

Article Snippet: More than one study found that 100 μM BzATP (Topscience # TP2226, Shanghai) can significantly activate P2X7 and change the cell membrane potential, so we also used the same concentration (Coddou et al., ).

Techniques: Activation Assay, De-Phosphorylation Assay, Western Blot, Phospho-proteomics, Analysis, Cell Culture, MANN-WHITNEY, Flow Cytometry, Mutagenesis, Negative Control, FACS, Immunofluorescence

Journal: Cell reports

Article Title: Single-cell transcriptomics of NRAS-mutated melanoma transitioning to drug resistance reveals P2RX7 as an indicator of early drug response.

doi: 10.1016/j.celrep.2023.112696

Figure Lengend Snippet: Figure 7. The impact of drug exposure on ROS production in NRASmut melanoma (A) ROS production after short-term (1 and 4 days) exposure to combined MEKi/CDK4/6i therapy in MelJuso (SAC) and SKMel30 (FAC) cells. Cell exposure to H2O2 for 30 min served as a positive control. Mean values ± SD from three biological replicates are plotted. (B) ROS production following P2RX7 silencing and/or activation with 100 mM BzATP in untreated or 4 days binimetinib/palbociclib-treated (B/P) MelJuso cells. RNAiMax, transfection reagent alone; NC, negative control siRNA. Bars indicate the mean values ± SD from three biological replicates. (C) Growth curves of SACs (MelJuso) during MEK/ CDK4/6 co-inhibition alone or with P2RX7 activation (100mMBzATP) and/orP2RX7antagonist A-740003 (1 mM), measured in an IncuCyte ZOOM live-cell microscope (representative results of mean of two technical replicates ±SEM). (D) ROS production after 4, 7, and 10 days of MEKi (binimetinib [B]) and CDK4/6i (palbociclib [P]) treat- ment (B/P) with or without BzATP (100 mM) and A-740003 (1 mM). For (A) and (B), statistical signifi- cance was established by one-way ANOVA, followed by multiple comparisons test; ns, not sig- nificant; *p % 0.05; **p % 0.01; ***p % 0.001; ****p % 0.0001.

Article Snippet: The P2RX7 agonist BzATP triethylammonium salt was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and reconstituted in water.

Techniques: Positive Control, Activation Assay, Transfection, Negative Control, Inhibition, Microscopy

Journal: Nature Communications

Article Title: Neutrophil P2X 7 receptors mediate NLRP3 inflammasome-dependent IL-1β secretion in response to ATP

doi: 10.1038/ncomms10555

Figure Lengend Snippet: ( a , b ). IL-1β in the cell supernatants of C57BL/6 bone-marrow-derived neutrophils primed 3 h with LPS (500 ng ml −1 ) and stimulated 45 min with ATP (0.5–3.0 mM) or 10 μM nigericin. Apyrase (10U ml −1 ) was added 30 min before stimulation with ATP or nigericin, and IL-1β secretion was quantified by ELISA. ( c ). IL-1β production by LPS-primed C57BL/6 murine neutrophils stimulated 45 min with BzATP. ( d ). IL-1β in supernatants of LPS-primed bone-marrow-derived neutrophils from C57BL/6, Nlrp3 −/− , Asc −/− and Caspase1,11 −/− mice after 45 min stimulation with 3 mM ATP or 10 μM nigericin. ( e , f ). Pro- and cleaved forms of caspase-1 and IL-1β in total cell lysates and TCA precipitated supernatant of LPS-primed C57BL/6 neutrophils after 45 min stimulation with 3 mM ATP or 10 μM nigericin. ( g ). LDH release as a measure of cytotoxicity in LPS-primed C57BL/6 neutrophils after ATP stimulation. ( h ). Propidium iodide (PI) uptake by bone marrow neutrophils and bone-marrow-derived macrophages following LPS priming and 45 min ATP (3 mM) stimulation. Data were generated by flow cytometry. Data points are mean±s.d. of at least four replicates per treatment and are representative of three independent experiments.

Article Snippet: BzATP triethylammonium salt (Tocris Bioscience) was dissolved in sterile water and used at millimolar concentrations.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Generated, Flow Cytometry

Journal: Frontiers in Cell and Developmental Biology

Article Title: Activation of P2×7 Receptor Promotes the Invasion and Migration of Colon Cancer Cells via the STAT3 Signaling

doi: 10.3389/fcell.2020.586555

Figure Lengend Snippet: Characterization of ATP-gated P2 × 7 receptor functionality in colon cancer cell lines. (A,B) : LOVO and SW480 cells were treated with ATP (0 μM), ATP (300 μM), BzATP (10 μM), ATP + A438079 (10 μM) or ATP + AZD9056 (10 μM), inward current of electrophysiological recording. (C,D) : LOVO and SW480 cells were treated with ATP (300 μM), BzATP (10 μM), ATP + A438079 (10 μM) or ATP + AZD9056 (10 μM). Fluo-4AM fluorescence technology was performed to detect calcium changes in colon cancer cells. The data came from the average of three independent experiments. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Cells were treated with or without P2 × 7 receptor agonist (ATP or BzATP), P2 × 7 receptor antagonist A438079 or AZD9056, or ATP + P2 × 7 receptor antagonist (A438019 or AZD9056) [These drugs are purchased from Glpbio (California, United States)], observed and photographed under an inverted microscope at 0 and 24 h, respectively, and the percentage of wound healing between cell scratches was calculated.

Techniques: Fluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: Activation of P2×7 Receptor Promotes the Invasion and Migration of Colon Cancer Cells via the STAT3 Signaling

doi: 10.3389/fcell.2020.586555

Figure Lengend Snippet: Activation of P2 × 7 receptor promoted the proliferation of colon cancer cells. (A–D) : LOVO and SW480 cells were treated with ATP (300 μM), BzATP (10 μM), A438079 (10 μM), AZD9056 (10 μM), ATP + A438079 (10 μM) or ATP + AZD9056 (10 μM) for 24 h. Edu assay was performed to detect the activity of colon cancer cells, bar = 100 μm, n = 3 independent experiments. ∗∗ P < 0.05, ∗∗∗ P < 0.01

Article Snippet: Cells were treated with or without P2 × 7 receptor agonist (ATP or BzATP), P2 × 7 receptor antagonist A438079 or AZD9056, or ATP + P2 × 7 receptor antagonist (A438019 or AZD9056) [These drugs are purchased from Glpbio (California, United States)], observed and photographed under an inverted microscope at 0 and 24 h, respectively, and the percentage of wound healing between cell scratches was calculated.

Techniques: Activation Assay, EdU Assay, Activity Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Activation of P2×7 Receptor Promotes the Invasion and Migration of Colon Cancer Cells via the STAT3 Signaling

doi: 10.3389/fcell.2020.586555

Figure Lengend Snippet: Cell scratch assay was performed to detect the role of extracellular ATP-medicated P2 × 7 receptor on the migration ability of colon cancer cells. (A,B) : LOVO and SW480 cells were treated with ATP (100–300 μM) or BzATP (10, 50, and 100 μM) for 24 h, and cell scratch assay was performed to detect the migration ability of colon cancer cells (bar = 50 μm). (C,D) : LOVO and SW480 cells were treated with ATP (300 μM), A438079 (10 μM), ATP + A438079 (10 μM), AZD9056 (10 μM) or ATP + AZD9056 (10 μM) for 24 h, and cell scratch assay was performed to detect the migration ability of colon cancer cells (bar = 50 μm) ( n = 3 independent experiments). ∗ P < 0.05, ∗∗ < 0.01.

Article Snippet: Cells were treated with or without P2 × 7 receptor agonist (ATP or BzATP), P2 × 7 receptor antagonist A438079 or AZD9056, or ATP + P2 × 7 receptor antagonist (A438019 or AZD9056) [These drugs are purchased from Glpbio (California, United States)], observed and photographed under an inverted microscope at 0 and 24 h, respectively, and the percentage of wound healing between cell scratches was calculated.

Techniques: Wound Healing Assay, Migration

Journal: Frontiers in Cell and Developmental Biology

Article Title: Activation of P2×7 Receptor Promotes the Invasion and Migration of Colon Cancer Cells via the STAT3 Signaling

doi: 10.3389/fcell.2020.586555

Figure Lengend Snippet: Role of P2 × 7 receptor in mediating the migration and invasion of colon cancer cells. (A–D) : LOVO and SW480 cells were treated with ATP (300 μM), A438079 (10 μM) or ATP + A438079 (10 μM) for 24 h. Effect of P2 × 7 receptor on the invasion and migration of LOVO and SW480 cells (bar = 100 μm). n = 3 independent experiments. ∗ P < 0.05 ATP VS Control; # P < 0.05 ATP + A438079 VS ATP.

Article Snippet: Cells were treated with or without P2 × 7 receptor agonist (ATP or BzATP), P2 × 7 receptor antagonist A438079 or AZD9056, or ATP + P2 × 7 receptor antagonist (A438019 or AZD9056) [These drugs are purchased from Glpbio (California, United States)], observed and photographed under an inverted microscope at 0 and 24 h, respectively, and the percentage of wound healing between cell scratches was calculated.

Techniques: Migration, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Activation of P2×7 Receptor Promotes the Invasion and Migration of Colon Cancer Cells via the STAT3 Signaling

doi: 10.3389/fcell.2020.586555

Figure Lengend Snippet: P2 × 7 receptor was involved in ATP-mediated the invasion and migration of colon cancer cells. (A) : RT-PCR was used to detect the expression of P2 × 7 receptor in LOVO and SW480 cells. (B,C) : LOVO and SW480 cells were transfected with control siRNA (siCtrl) or P2 × 7 siRNA (siP2 × 7) for 24 h, and knockdown efficiency was examined by real-time PCR and Western-blotting. ∗ P < 0.05 siP2 × 7 VS Control. (D,E) : Effect of P2 × 7 receptor knockdown on ATP-induced the invasion and migration of LOVO and SW480 cells. n = 3 independent experiments. ∗ P < 0.05 siCtrl ATP VS siCtrl control; # P < 0.05 siP2 × 7 ATP VS siCtrl ATP.

Article Snippet: Cells were treated with or without P2 × 7 receptor agonist (ATP or BzATP), P2 × 7 receptor antagonist A438079 or AZD9056, or ATP + P2 × 7 receptor antagonist (A438019 or AZD9056) [These drugs are purchased from Glpbio (California, United States)], observed and photographed under an inverted microscope at 0 and 24 h, respectively, and the percentage of wound healing between cell scratches was calculated.

Techniques: Migration, Reverse Transcription Polymerase Chain Reaction, Expressing, Transfection, Control, Knockdown, Real-time Polymerase Chain Reaction, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Activation of P2×7 Receptor Promotes the Invasion and Migration of Colon Cancer Cells via the STAT3 Signaling

doi: 10.3389/fcell.2020.586555

Figure Lengend Snippet: Effect of P2 × 7 receptor on the expression of E-cadherin and MMP-2. (A,B) : LOVO and SW480 cells were treated with ATP (300 μM) or BzATP (10 μM), Western-blotting was used to analysis the protein expression of E-cadherin. (C,D) : LOVO and SW480 cells were treated with ATP (300 μM) or BzATP (10 μM), real-time PCR and ELISA were used to analysis the expression level of MMP-2 mRNA and the concentration level of MMP-2. n = 3 independent experiments. ∗ P < 0.05 ATP VS Control; # P < 0.05 BzATP VS Control.

Article Snippet: Cells were treated with or without P2 × 7 receptor agonist (ATP or BzATP), P2 × 7 receptor antagonist A438079 or AZD9056, or ATP + P2 × 7 receptor antagonist (A438019 or AZD9056) [These drugs are purchased from Glpbio (California, United States)], observed and photographed under an inverted microscope at 0 and 24 h, respectively, and the percentage of wound healing between cell scratches was calculated.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Concentration Assay, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Activation of P2×7 Receptor Promotes the Invasion and Migration of Colon Cancer Cells via the STAT3 Signaling

doi: 10.3389/fcell.2020.586555

Figure Lengend Snippet: Effect of P2 × 7 receptor knockdown on the expression of E-cadherin, MMP-2, N-cadherin, Vementin, Zeb1 and Snail (A,B) : Western-blotting was used to analysis of the expression level of E-cadherin in siCtrl or siP2 × 7 LOVO and SW480 cells. (C,D) : Real-time PCR and ELISA were used to detect the expression of MMP-2 in siCtrl or siP2 × 7 LOVO and SW480 cells. (E,F) : Western-blotting and real-time PCR was performed to measure the expression of N-cadherin, Vimentin, Zeb1 and Snail preotein and mRNA in siCtrl or siP2 × 7 LOVO and SW480 cells. n = 3 independent experiments. ∗ P < 0.05 siCtrl ATP VS siCtrl con; ∗ P < 0.05 siCtrl ATP VS siCtrl con. ∗∗ P < 0.05 siP2 × 7 ATP VS siCtrl ATP, # P < 0.05 siP2 × 7 ATP VS siP2 × 7 con.

Article Snippet: Cells were treated with or without P2 × 7 receptor agonist (ATP or BzATP), P2 × 7 receptor antagonist A438079 or AZD9056, or ATP + P2 × 7 receptor antagonist (A438019 or AZD9056) [These drugs are purchased from Glpbio (California, United States)], observed and photographed under an inverted microscope at 0 and 24 h, respectively, and the percentage of wound healing between cell scratches was calculated.

Techniques: Knockdown, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Activation of P2×7 Receptor Promotes the Invasion and Migration of Colon Cancer Cells via the STAT3 Signaling

doi: 10.3389/fcell.2020.586555

Figure Lengend Snippet: ATP-mediated activation of P2 × 7 receptor activated STAT3 signaling. (A,B): ATP (300 μM) stimulated LOVO and SW480 cells at 0, 30, 60, 90, and 120 min. Western-blotting was used to detect the expression level of p-STAT3. (C,D): LOVO and SW480 cells were treated with siCtrl or siP2 × 7 for 90 min. Western blotting was performed to detect the expression of p-STAT3 protein. n = 3 independent experiments. ∗ P < 0.05 siCtrl ATP VS siCtrl con; ∗∗ P < 0.05 siP2 × 7 ATP VS siCtrl ATP; # P < 0.05 siP2 × 7 ATP VS siP2 × 7 con.

Article Snippet: Cells were treated with or without P2 × 7 receptor agonist (ATP or BzATP), P2 × 7 receptor antagonist A438079 or AZD9056, or ATP + P2 × 7 receptor antagonist (A438019 or AZD9056) [These drugs are purchased from Glpbio (California, United States)], observed and photographed under an inverted microscope at 0 and 24 h, respectively, and the percentage of wound healing between cell scratches was calculated.

Techniques: Activation Assay, Western Blot, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Activation of P2×7 Receptor Promotes the Invasion and Migration of Colon Cancer Cells via the STAT3 Signaling

doi: 10.3389/fcell.2020.586555

Figure Lengend Snippet: The role of P2 × 7 receptor mediated STAT3 in the migration and invasion of colon cancer cells. (A,B) : On the basis of ATP, LOVO cells were treated with STA-21 (10, 30, and 50 μM) (ATP + STA-21), respectively, and detected the migration and invasion abilities of LOVO cells by transwell assay. (C,D) : SW480 cells were treated with ATP + STA-21 (10, 30, and 50 μM), respectively, and detected the migration and invasion abilities of SW480 cells by transwell assay. n = 3 independent experiments. ∗ P < 0.05 ATP VS Con; ∗∗ P < 0.05 STA-21 VS ATP; # P < 0.05 compared between STA-21.

Article Snippet: Cells were treated with or without P2 × 7 receptor agonist (ATP or BzATP), P2 × 7 receptor antagonist A438079 or AZD9056, or ATP + P2 × 7 receptor antagonist (A438019 or AZD9056) [These drugs are purchased from Glpbio (California, United States)], observed and photographed under an inverted microscope at 0 and 24 h, respectively, and the percentage of wound healing between cell scratches was calculated.

Techniques: Migration, Transwell Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Activation of P2×7 Receptor Promotes the Invasion and Migration of Colon Cancer Cells via the STAT3 Signaling

doi: 10.3389/fcell.2020.586555

Figure Lengend Snippet: Effect of STAT3 pathway on the expression of E-cadherin and MMP-2 in colon cancer cells mediated by P2 × 7 receptor. LOVO and SW480 cells were treated with ATP (300 μM) or ATP + STA-21 (30 and 50 μM). (A,B) : Western-blotting was performed to analysis of protein levels of E-cadherin in LOVO and SW480 cells. ( C,D) : Real-time PCR and ELISA were used to analysis of MMP-2 expression. n = 3 independent experiments. ∗ P < 0.05 ATP VS Con; # P < 0.05 STA-21 VS ATP.

Article Snippet: Cells were treated with or without P2 × 7 receptor agonist (ATP or BzATP), P2 × 7 receptor antagonist A438079 or AZD9056, or ATP + P2 × 7 receptor antagonist (A438019 or AZD9056) [These drugs are purchased from Glpbio (California, United States)], observed and photographed under an inverted microscope at 0 and 24 h, respectively, and the percentage of wound healing between cell scratches was calculated.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay