Journal: Journal of Neuroinflammation

Article Title: P2X7R blockade prevents NLRP3 inflammasome activation and brain injury in a rat model of intracerebral hemorrhage: involvement of peroxynitrite

doi: 10.1186/s12974-015-0409-2

Figure Lengend Snippet: Expression profile of P2X7R and its cellular location after collagenase-induced intracerebral hemorrhage (ICH). Western blot analysis ( a ) for the time course of P2X7R expression in the ipsilateral hemisphere of Sham and ICH rats within 72 h. Quantification of P2X7R ( b ) expression is shown, n = 4 rats per group and time point. Confocal images ( c ) of double immunofluorescence for P2X7R expression in Iba-1-positive microglia 24 h following ICH, n = 6 rats per group. Scale bar = 12.5 μm. Data represent means ± SD. * P < 0.05 vs. Sham, ** P < 0.01 vs. Sham. GAPDH glyceraldehyde 3-phosphate dehydrogenase, ICH intracerebral hemorrhage

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-P2X7R (1:1000, Alomone Labs, Jerusalem, Israel), rabbit polyclonal anti-NLRP3 antibody (1:1000, Santa Cruz, Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-ASC antibody (1:500, Abclonal, Cambridge, MA, USA), mouse monoclonal anti-caspase-1 p20 antibody (1:1000, Santa Cruz Biotechnology), rabbit polyclonal anti-IL-1β antibody (1:1000, Millipore), rabbit monoclonal anti-IL-18 antibody (1:1000, Abcam, Cambridge, UK), mouse monoclonal anti-nitrotyrosine antibody (1:1000, Abcam), mouse monoclonal anti-iNOS antibody (1:200, Santa Cruz Biotechnology), mouse monoclonal anti-gp91 phox antibody (1:2000, BD Transduction Laboratories, San Jose, CA, USA), and rabbit polyclonal anti-myeloperoxidase antibody (MPO, 1:500, Abcam).

Techniques: Expressing, Western Blot, Immunofluorescence

Journal: Journal of Neuroinflammation

Article Title: P2X7R blockade prevents NLRP3 inflammasome activation and brain injury in a rat model of intracerebral hemorrhage: involvement of peroxynitrite

doi: 10.1186/s12974-015-0409-2

Figure Lengend Snippet: Effects of P2X7R small interfering RNA (siRNA) treatment in ICH rats. RT-PCR of P2X7R after siRNA treatment 24 h following ICH, n = 6 rats per group ( a ). Western blot assay and quantification of P2X7R ( b ) after siRNA treatment 24 h following ICH, n = 4 rats per group. Brain edema ( c ) at 24 h after ICH, n = 6 rats per group. mNSS ( d ) at 24 and 72 h after ICH, n = 6 rats per group. Data represent means ± SD. ** P < 0.01 vs. Sham, @ P < 0.05 vs. Vehicle, # P < 0.05 vs. ICH + Scramble siRNA. GAPDH glyceraldehyde 3-phosphate dehydrogenase, mNSS modified Neurological Severity Score, siRNA small interfering RNA

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-P2X7R (1:1000, Alomone Labs, Jerusalem, Israel), rabbit polyclonal anti-NLRP3 antibody (1:1000, Santa Cruz, Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-ASC antibody (1:500, Abclonal, Cambridge, MA, USA), mouse monoclonal anti-caspase-1 p20 antibody (1:1000, Santa Cruz Biotechnology), rabbit polyclonal anti-IL-1β antibody (1:1000, Millipore), rabbit monoclonal anti-IL-18 antibody (1:1000, Abcam, Cambridge, UK), mouse monoclonal anti-nitrotyrosine antibody (1:1000, Abcam), mouse monoclonal anti-iNOS antibody (1:200, Santa Cruz Biotechnology), mouse monoclonal anti-gp91 phox antibody (1:2000, BD Transduction Laboratories, San Jose, CA, USA), and rabbit polyclonal anti-myeloperoxidase antibody (MPO, 1:500, Abcam).

Techniques: Small Interfering RNA, Reverse Transcription Polymerase Chain Reaction, Western Blot, Modification

Journal: Journal of Neuroinflammation

Article Title: P2X7R blockade prevents NLRP3 inflammasome activation and brain injury in a rat model of intracerebral hemorrhage: involvement of peroxynitrite

doi: 10.1186/s12974-015-0409-2

Figure Lengend Snippet: Effects of P2X7R siRNA on NLRP3 inflammasome activation and IL-1β/IL-18 maturation after ICH. RT-PCR of NLRP3 after P2X7R siRNA treatment 24 h following ICH ( a ), n = 6 rats per group. Western blot assay ( b ) and quantification of NLRP3 ( c ), ASC ( d ), caspase-1 p20 subunit ( e ), IL-1β ( f ), and IL-18 ( g ) after P2X7R siRNA treatment 24 h following ICH, n = 4 rats per group. Data represent means ± SD. ** P < 0.01 vs. Sham, @ P < 0.05 vs. Vehicle, # P < 0.05 vs. ICH + Scramble siRNA. ASC adaptor protein apoptosis-associated speck-like protein containing a CARD, GAPDH glyceraldehyde 3-phosphate dehydrogenase, ICH intracerebral hemorrhage, IL interleukin, NLRP3 The NLR family, pyrin domain-containing 3

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-P2X7R (1:1000, Alomone Labs, Jerusalem, Israel), rabbit polyclonal anti-NLRP3 antibody (1:1000, Santa Cruz, Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-ASC antibody (1:500, Abclonal, Cambridge, MA, USA), mouse monoclonal anti-caspase-1 p20 antibody (1:1000, Santa Cruz Biotechnology), rabbit polyclonal anti-IL-1β antibody (1:1000, Millipore), rabbit monoclonal anti-IL-18 antibody (1:1000, Abcam, Cambridge, UK), mouse monoclonal anti-nitrotyrosine antibody (1:1000, Abcam), mouse monoclonal anti-iNOS antibody (1:200, Santa Cruz Biotechnology), mouse monoclonal anti-gp91 phox antibody (1:2000, BD Transduction Laboratories, San Jose, CA, USA), and rabbit polyclonal anti-myeloperoxidase antibody (MPO, 1:500, Abcam).

Techniques: Activation Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Journal of Neuroinflammation

Article Title: P2X7R blockade prevents NLRP3 inflammasome activation and brain injury in a rat model of intracerebral hemorrhage: involvement of peroxynitrite

doi: 10.1186/s12974-015-0409-2

Figure Lengend Snippet: Effects of BBG on NLRP3 inflammasome activation and IL-1β/IL-18 maturation. RT-PCR of NLRP3 after BBG treatment 24 h following ICH, n = 6 rats per group ( a ). Representative western blot ( b ) and therapeutic effects of BBG on P2X7R ( c ), NLRP3 ( d ), ASC ( e ), caspase-1 p20 subunit ( f ), mature IL-1β ( g ), and mature IL-18 ( h ) levels in the ipsilateral hemisphere 24 h after ICH, n = 4 rats per group. Data represent means ± SD. * P < 0.05, ** P < 0.01. ASC adaptor protein apoptosis-associated speck-like protein containing a CARD, BBG brilliant blue G, GAPDH glyceraldehyde 3-phosphate dehydrogenase, ICH intracerebral hemorrhage, IL interleukin, NLRP3 pyrin domain-containing 3

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-P2X7R (1:1000, Alomone Labs, Jerusalem, Israel), rabbit polyclonal anti-NLRP3 antibody (1:1000, Santa Cruz, Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-ASC antibody (1:500, Abclonal, Cambridge, MA, USA), mouse monoclonal anti-caspase-1 p20 antibody (1:1000, Santa Cruz Biotechnology), rabbit polyclonal anti-IL-1β antibody (1:1000, Millipore), rabbit monoclonal anti-IL-18 antibody (1:1000, Abcam, Cambridge, UK), mouse monoclonal anti-nitrotyrosine antibody (1:1000, Abcam), mouse monoclonal anti-iNOS antibody (1:200, Santa Cruz Biotechnology), mouse monoclonal anti-gp91 phox antibody (1:2000, BD Transduction Laboratories, San Jose, CA, USA), and rabbit polyclonal anti-myeloperoxidase antibody (MPO, 1:500, Abcam).

Techniques: Activation Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: The Journal of Physiology

Article Title: Stretch‐induced Ca 2+ independent ATP release in hippocampal astrocytes

doi: 10.1113/JP275805

Figure Lengend Snippet: A, images of representative astrocytes double‐labelled with antibodies against GFAP (red) and P2X7 receptors (green). Scale bar = 20 μm. B, BBG (100 nm), a specific P2X7 receptor antagonist, inhibited MARA in astrocytes. Inset: BBG (100 nm) did not affect the sensitivity of ATP sniffer currents (n = 6). C, summary of the effect of BBG on evoked ATP release (n = 6). ** P < 0.01. D, immunofluorescence staining showing partial knockdown of P2X7 receptors. Scale bar = 20 μm. E, upper, western blots showing partial knockdown of P2X7 receptors by RNAi. Lower: normalized P2X7 protein levels were 100% (control), 93 ± 9% (scrambled) and 39 ± 13% (RNAi) (n = 9). ** P < 0.01. F, MARA was markedly lower in P2X7 knockdown astrocytes than in normal astrocytes, or scrambled RNAi‐transfected astrocytes. G, summary of the RNAi knockdown effect on MARA (n = 20). *** P < 0.001.

Article Snippet: P2X7 short hairpin (sh)RNA(r) and scrambled lentiviral particles were obtained from Santa Cruz Biotechnology.

Techniques: Immunofluorescence, Staining, Knockdown, Western Blot, Control, Transfection

Journal: The Journal of Physiology

Article Title: Stretch‐induced Ca 2+ independent ATP release in hippocampal astrocytes

doi: 10.1113/JP275805

Figure Lengend Snippet: A and B, RT‐PCR for mRNA of P2X7 receptors and pannexin‐1 in cultured astrocytes (A) and C6 cells (B). C, comparison of two normalized typical MARA recordings in an astrocyte (grey trace) and a pannexin‐1‐null glioma C6 cell (black trace). D, statistical comparisons of half‐widths and rise‐times of cultured astrocytes (n = 78) and C6 cells (n = 12).

Article Snippet: P2X7 short hairpin (sh)RNA(r) and scrambled lentiviral particles were obtained from Santa Cruz Biotechnology.

Techniques: Reverse Transcription Polymerase Chain Reaction, Cell Culture, Comparison

Journal: The Journal of Physiology

Article Title: Stretch‐induced Ca 2+ independent ATP release in hippocampal astrocytes

doi: 10.1113/JP275805

Figure Lengend Snippet: A, expression of P2X7 channels in astrocytes in hippocampal slices. Upper: hippocampal slice from a P2X7‐EGFP transgenic mouse immunostained for GFAP (red) to identify astrocytes; the nuclei were stained with DAPI (blue). Scale bar = 250 μm. Middle: magnified image of boxed area in upper panel. Scale bar = 50 μm. Lower: magnified view of boxed area in middle panel reveals that P2X7 and GFAP co‐localized in astrocytes. Scale bar = 10 μm. B, hippocampal slice from a P2X7‐EGFP transgenic mouse loaded with Fluo4 (10 μm, white) and SR101 (0.5 μm, red). C, left: ATP‐induced Ca2+ elevation in a P2X7‐positive astrocyte was blocked by BBG (200 nm). Right: summary of the BBG effect in P2X7‐positive astrocytes (n = 15). D, left: ATP‐induced Ca2+ elevation in a P2X7‐negative cell was not blocked by BBG (200 nm). Right: summary of the BBG effect in P2X7‐negative cells (n = 6).

Article Snippet: P2X7 short hairpin (sh)RNA(r) and scrambled lentiviral particles were obtained from Santa Cruz Biotechnology.

Techniques: Expressing, Transgenic Assay, Staining

Journal: Molecular Therapy Oncolytics

Article Title: Cathelicidin Suppresses Colon Cancer Metastasis via a P2RX7-Dependent Mechanism

doi: 10.1016/j.omto.2019.01.004

Figure Lengend Snippet: Cathelicidin-Mediated Inhibition of Colon Cancer Cell Migration Was P2RX7 Dependent (A) SW620 cells were transfected with control lentivirus or TUBB3-overexpressing lentivirus, followed by exposure to LL-37. Cell migration of SW620 cells. (B) SW620 cells were transiently transfected with control small interfering RNA (siRNA) or P2RX7 siRNA (80 pmol/mL), followed by exposure to LL-37. TUBB3 mRNA expression. (C) CAMP, (D) FPRL1, and (E) P2RX7 mRNA expression in human colon cancer PCR array plate. (F) Cell migration of SW620 cells. SW620 cells were treated with DMSO, KN62, and WRW4 for 30 min, followed by LL-37 for 7 h. Results were pooled from three independent experiments.

Article Snippet: Some of the nude mice were injected intraperitoneally with P2RX7 antagonist KN62 (5 mg/kg, Tocris) or control DMSO solution (50 μL per mouse) daily from day 21 to day 28.

Techniques: Inhibition, Migration, Transfection, Control, Small Interfering RNA, Expressing

Journal: Molecular Therapy Oncolytics

Article Title: Cathelicidin Suppresses Colon Cancer Metastasis via a P2RX7-Dependent Mechanism

doi: 10.1016/j.omto.2019.01.004

Figure Lengend Snippet: Cathelicidin Inhibited TUBB3 Expression via P2RX7 (A) Green tubulin tracker staining with blue nuclear staining in human cancer SW620 cells. SW620 cells were pretreated with DMSO (10 μL/mL) or KN62 (10 μM) for 30 min, followed by exposure to LL-37 (5 μM) for 24 h. LL-37 reduced tubulin expression in SW620 cells that was prevented by KN62. (B) SW620 cells were transiently transfected with control small interfering RNA (siRNA) or P2RX7 shRNA (1 μg/mL), followed by exposure to LL-37. TUBB3 mRNA expression in SW620 cells. (C) P2RX7 mRNA expression in the transfected SW620 cells. (D) HT-29 cells were pretreated with DMSO (10 μL/mL) or KN62 (10 μM) for 30 min, followed by exposure to LL-37. TUBB3 mRNA expression. Results were pooled from three independent experiments.

Article Snippet: Some of the nude mice were injected intraperitoneally with P2RX7 antagonist KN62 (5 mg/kg, Tocris) or control DMSO solution (50 μL per mouse) daily from day 21 to day 28.

Techniques: Expressing, Staining, Transfection, Control, Small Interfering RNA, shRNA

Journal: Molecular Therapy Oncolytics

Article Title: Cathelicidin Suppresses Colon Cancer Metastasis via a P2RX7-Dependent Mechanism

doi: 10.1016/j.omto.2019.01.004

Figure Lengend Snippet: Cathelicidin Inhibited Colon Cancer Metastasis via P2RX7 (A and B) Human cytokeratin-18 expression (representing human colon cancer cells) in (A) lungs and (B) liver of nude mice was identified by brown color spots (indicated by arrows). Intravenous cathelicidin expressing AAV administration reduced human CK18 expression in lungs and liver of nude mice.

Article Snippet: Some of the nude mice were injected intraperitoneally with P2RX7 antagonist KN62 (5 mg/kg, Tocris) or control DMSO solution (50 μL per mouse) daily from day 21 to day 28.

Techniques: Expressing

Journal: Molecular Therapy Oncolytics

Article Title: Cathelicidin Suppresses Colon Cancer Metastasis via a P2RX7-Dependent Mechanism

doi: 10.1016/j.omto.2019.01.004

Figure Lengend Snippet: Cathelicidin Inhibited TUBB3 mRNA Expression in Metastasized Tumors via P2RX7 (A and B) Human keratin 20 mRNA expression (human colon cancer cell marker) in (A) lungs and (B) liver of nude mice was significantly reduced by CAMP -HA-AAV. Intraperitoneal KN62 treatment increased the presence of cytokeratin 18 protein and keratin 20 mRNA expression in the CAMP -HA-AAV-treated nude mice. (C and D) Human TUBB3 mRNA expression in (C) lungs and (D) liver of nude mice was significantly reduced by CAMP -HA-AAV. Intraperitoneal KN62 treatment increased the presence of human TUBB3 mRNA expression in the CAMP -HA-AAV-treated nude mice. (E) Diameters of subcutaneous tumors. Intravenous CAMP -HA-AAV significantly reduced subcutaneous tumor diameters in nude mice that were partially reversed by KN62 treatment.

Article Snippet: Some of the nude mice were injected intraperitoneally with P2RX7 antagonist KN62 (5 mg/kg, Tocris) or control DMSO solution (50 μL per mouse) daily from day 21 to day 28.

Techniques: Expressing, Marker

Journal: Molecular Therapy Oncolytics

Article Title: Cathelicidin Suppresses Colon Cancer Metastasis via a P2RX7-Dependent Mechanism

doi: 10.1016/j.omto.2019.01.004

Figure Lengend Snippet: Primers for Real-Time RT-PCR

Article Snippet: Some of the nude mice were injected intraperitoneally with P2RX7 antagonist KN62 (5 mg/kg, Tocris) or control DMSO solution (50 μL per mouse) daily from day 21 to day 28.

Techniques:

Journal: Basic Research in Cardiology

Article Title: Renal denervation attenuates cardiac dysfunction in HFpEF by inhibiting the ATP-P2X7-NLRP3 inflammasome axis

doi: 10.1007/s00395-025-01138-5

Figure Lengend Snippet: Renal denervation reduces NLRP3 inflammasome activation induced by the ATP-P2X7 axis. a ATP levels in heart lysates (n = 6). b P2X7 mRNA expression in heart tissue, determined by qRT-PCR (n = 6). c , d Western blot analysis of P2X7, NLRP3 ( c ), ASC, and cleaved-caspase-1 ( d ) in mouse hearts. e Quantification of protein expression levels from ( c , d ) (n = 3). f Immunohistochemical staining of NLRP3 in myocardial sections. g Semi-quantitative scoring of NLRP3 immunoreactivity: 0 (negative), 1 (weak), 2 (mild), 3 (moderate), 4 (strong) (n = 6). h NLRP3 immunofluorescence in H9c2 cardiomyocytes transfected with control siRNA (si-Ctrl) or P2X7 siRNA (si-P2X7) and stimulated with ATP (5 mM). Right panel: relative mean fluorescence intensity (MFI) quantification (n = 6). i , j Western blot of P2X7, NLRP3 ( i ), ASC, and cleaved-caspase-1 ( j ) in H9c2 cells under the same treatments. k Quantification of the protein levels shown in ( i , j ) (n = 3). l Schematic of RNA-seq workflow in neonatal rat ventricular myocytes (NRVMs): Control, ATP, and ATP plus the P2X7 antagonist A438079. m Volcano plot of differentially expressed genes for ATP vs. Control; selected genes related to NLRP3 inflammasome/NF-κB/leukocyte activation are highlighted (FDR-adjusted P shown on the y-axis). n Integrated transcriptomics: Venn diagram showing ATP-induced DEGs reversed by A438079, with a heat map of representative shared genes across groups. One-way ANOVA with Tukey’s post hoc test for ( a, b, e, h and k ); Kruskal–Wallis with Dunn’s post hoc for the ordinal IHC score ( g ). Omics significance was assessed by FDR-adjusted P values (Benjamini-Hochberg). Data are mean ± SD unless noted. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Proteins were detected with the following primary antibodies: P2X7 (11144–1-AP, Proteintech, Rosemont, IL, USA); NLRP3 (30109–1-AP, Proteintech, Rosemont, IL, USA); apoptosis-associated speck-like protein containing a CARD (ASC) (30641–1-AP, Proteintech, Rosemont, IL, USA); Cleaved-Caspase1 (89332, Cell Signaling Technology, Danvers, MA, USA); Caspase1 (83383, Cell Signaling Technology, Danvers, MA, USA); catalase (CAT) (21260–1-AP, Proteintech, Rosemont, IL, USA); glutathione peroxidase 4 (GPX4) (67763–1-Ig, Proteintech, Rosemont, IL, USA); superoxide dismutase 2, mitochondrial (SOD2) (24127–1-AP, Proteintech, Rosemont, IL, USA).

Techniques: Activation Assay, Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemical staining, Staining, Immunofluorescence, Transfection, Control, Fluorescence, RNA Sequencing

Journal: Basic Research in Cardiology

Article Title: Renal denervation attenuates cardiac dysfunction in HFpEF by inhibiting the ATP-P2X7-NLRP3 inflammasome axis

doi: 10.1007/s00395-025-01138-5

Figure Lengend Snippet: Pharmacologic and genetic inhibition of P2X7 mitigates ATP-evoked mitochondrial ROS, oxidative stress, and pyroptosis in vitro. Neonatal Sprague–Dawley rat cardiomyocytes (NRCMs) were pre-treated with the P2X7 antagonist A-438079 (10 µM, 1 h) and then challenged with ATP (5 mM) for 24 h; vehicle was 0.1% DMSO. An A-438079-alone group was included. H9c2 cardiomyocytes were transfected with control siRNA or P2rx7 (P2X7) siRNA and stimulated with ATP (5 mM, 24 h). a MitoTracker staining showing mitochondrial networks under the indicated NRCM treatments. b, c Quantification of mitochondrial network metrics in NRCMs: footprint (µm 2 ) ( b ) and individuals ( c ). d Representative MitoSOX staining (with DAPI) visualizing mitochondrial superoxide in NRCMs. e MitoSOX mean fluorescence intensity (MFI) in NRCMs (n = 6). f Representative DCFH-DA fluorescence images of intracellular ROS (green) with DAPI nuclear counterstain (blue) in H9c2 cardiomyocytes. g Quantification of DCFH-DA mean fluorescence intensity in H9c2 cardiomyocytes (MFI, fold to Control) (n = 3). h-j Oxidative-stress markers in NRCMs: 4-hydroxynonenal (4-HNE) ( h ), 8-OHdG ( i ), catalase (CAT) ( j ) (n = 4). l Western blots of key antioxidant enzymes (CAT, GPX4, SOD2) in H9c2 cells under siRNA treatments. l – o Cytotoxicity/pyroptosis readouts in NRCMs subjected to the pharmacologic protocol: LDH release ( l ), caspase-1 activity ( m ), and IL-1β ( n ) and IL-18 ( o ) in supernatants (n = 4). One-way ANOVA with Tukey’s post hoc was performed for statistical analysis. Data is presented as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: Proteins were detected with the following primary antibodies: P2X7 (11144–1-AP, Proteintech, Rosemont, IL, USA); NLRP3 (30109–1-AP, Proteintech, Rosemont, IL, USA); apoptosis-associated speck-like protein containing a CARD (ASC) (30641–1-AP, Proteintech, Rosemont, IL, USA); Cleaved-Caspase1 (89332, Cell Signaling Technology, Danvers, MA, USA); Caspase1 (83383, Cell Signaling Technology, Danvers, MA, USA); catalase (CAT) (21260–1-AP, Proteintech, Rosemont, IL, USA); glutathione peroxidase 4 (GPX4) (67763–1-Ig, Proteintech, Rosemont, IL, USA); superoxide dismutase 2, mitochondrial (SOD2) (24127–1-AP, Proteintech, Rosemont, IL, USA).

Techniques: Inhibition, In Vitro, Transfection, Control, Staining, Fluorescence, Western Blot, Activity Assay

Journal: Basic Research in Cardiology

Article Title: Renal denervation attenuates cardiac dysfunction in HFpEF by inhibiting the ATP-P2X7-NLRP3 inflammasome axis

doi: 10.1007/s00395-025-01138-5

Figure Lengend Snippet: Pharmacological inhibition of P2X7 by A438079 improves HFpEF phenotypes. a Schematic of the experimental design. After 3-hit HFpEF induction, mice received A438079 (5 mg/kg, via osmotic pump) or vehicle for two weeks. b WGA staining of myocardial sections showing cross-sectional area of cardiomyocytes, quantified on the right. c Masson’s trichrome staining to evaluate myocardial fibrosis, with fibrosis percentage shown on the right. d mRNA expression of Col1a2, Timp1, Fn1, and α-SMA in hearts. e Representative H&E staining of heart sections. f Representative images of CD31 immunohistochemical staining in heart sections. g Serum NT-proBNP. h Heart weight normalized to tibia length (HW/TL). i Treadmill running distance for exercise capacity. j Left ventricular ejection fraction measured by echocardiography. k Ratio of early Doppler transmitral flow velocity (E) to tissue Doppler velocity (e′). One-way ANOVA with Tukey’s post hoc was performed for statistical analysis. n = 6 mice/group. Data is presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Proteins were detected with the following primary antibodies: P2X7 (11144–1-AP, Proteintech, Rosemont, IL, USA); NLRP3 (30109–1-AP, Proteintech, Rosemont, IL, USA); apoptosis-associated speck-like protein containing a CARD (ASC) (30641–1-AP, Proteintech, Rosemont, IL, USA); Cleaved-Caspase1 (89332, Cell Signaling Technology, Danvers, MA, USA); Caspase1 (83383, Cell Signaling Technology, Danvers, MA, USA); catalase (CAT) (21260–1-AP, Proteintech, Rosemont, IL, USA); glutathione peroxidase 4 (GPX4) (67763–1-Ig, Proteintech, Rosemont, IL, USA); superoxide dismutase 2, mitochondrial (SOD2) (24127–1-AP, Proteintech, Rosemont, IL, USA).

Techniques: Inhibition, Staining, Expressing, Immunohistochemical staining

Journal: FEBS letters

Article Title: Low-intensity pulsed ultrasound-induced ATP increases bone formation via the P2X7 receptor in osteoblast-like MC3T3-E1 cells.

doi: 10.1016/j.febslet.2014.12.013

Figure Lengend Snippet: Fig. 2. Effects of LIPUS on the expression of RUNX2, Osterix, and the P2X7 receptor. MC3T3-E1 cells were stimulated with LIPUS (5, 10, 20, and 30 min) or left without stimulation (control) and mRNA expression of RUNX2 (a), Osterix (b), and P2X7 (c) was determined on days 1, 3 and 5 of culture using real-time PCR. Data are expressed as the mean ± S.E.M. of three independent experiments performed in triplicate; ⁄P < 0.05, ⁄⁄P < 0.01, LIPUS vs. control.

Article Snippet: To obtain stable P2X7-deficient clones, the cells were transfected with Lentiviral Particles containing the puromycin resistance gene for antibiotic selection and P2X7 short hairpin RNA (shRNA) or non-target (scrambled) control shRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and cultured in the presence of 5 lg/ml puromycin (Enzo Life Sciences Inc., Farmingdale, NY, USA).

Techniques: Expressing, Control, Real-time Polymerase Chain Reaction

Journal: FEBS letters

Article Title: Low-intensity pulsed ultrasound-induced ATP increases bone formation via the P2X7 receptor in osteoblast-like MC3T3-E1 cells.

doi: 10.1016/j.febslet.2014.12.013

Figure Lengend Snippet: Fig. 8. Effects of LIPUS on mRNA expression of ECM proteins in P2X7-deficient osteoblasts. MC3T3-E1 cells transfected with P2X7 shRNA (sh P2X7) or scrambled shRNA (sh control) were stimulated or not with LIPUS for 30 min per day during 14 days and mRNA expression of type I collagen (a), bone sialoprotein (b), osteopontin (c), and osteocalcin (d) was determined by real-time PCR on days 3, 7, and 14 of culture. Data are expressed as the mean ± S.E.M. of three independent experiments performed in triplicate; ⁄P < 0.05, ⁄⁄P < 0.01, sh control + LIPUS vs. sh control, +P < 0.05, ++P < 0.01, sh P2X7 + LIPUS vs. sh control + LIPUS.

Article Snippet: To obtain stable P2X7-deficient clones, the cells were transfected with Lentiviral Particles containing the puromycin resistance gene for antibiotic selection and P2X7 short hairpin RNA (shRNA) or non-target (scrambled) control shRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and cultured in the presence of 5 lg/ml puromycin (Enzo Life Sciences Inc., Farmingdale, NY, USA).

Techniques: Expressing, Transfection, shRNA, Control, Real-time Polymerase Chain Reaction

Journal: FEBS letters

Article Title: Low-intensity pulsed ultrasound-induced ATP increases bone formation via the P2X7 receptor in osteoblast-like MC3T3-E1 cells.

doi: 10.1016/j.febslet.2014.12.013

Figure Lengend Snippet: Fig. 7. Effects of P2X7-specific shRNA on P2X7 gene silencing in MC3T3-E1 cells. MC3T3-E1 cells were transfected with Lentiviral Particles containing P2X7-specific shRNA (shP2X7) or control scrambled shRNA (sh control). Total RNA and protein was purified from confluent cells, and the expression of P2X7 mRNA and protein was determined by real-time PCR (a) and Western blotting (b); histograms show the intensity of Western blotting bands under each condition (c). Data are expressed as the mean ± S.E.M. of three independent experiments performed in triplicate; ⁄P < 0.05, ⁄⁄P < 0.01, sh P2X7 vs.sh control.

Article Snippet: To obtain stable P2X7-deficient clones, the cells were transfected with Lentiviral Particles containing the puromycin resistance gene for antibiotic selection and P2X7 short hairpin RNA (shRNA) or non-target (scrambled) control shRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and cultured in the presence of 5 lg/ml puromycin (Enzo Life Sciences Inc., Farmingdale, NY, USA).

Techniques: shRNA, Transfection, Control, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Molecular Therapy Oncolytics

Article Title: Cathelicidin Suppresses Colon Cancer Metastasis via a P2RX7-Dependent Mechanism

doi: 10.1016/j.omto.2019.01.004

Figure Lengend Snippet: Cathelicidin-Mediated Inhibition of Colon Cancer Cell Migration Was P2RX7 Dependent (A) SW620 cells were transfected with control lentivirus or TUBB3-overexpressing lentivirus, followed by exposure to LL-37. Cell migration of SW620 cells. (B) SW620 cells were transiently transfected with control small interfering RNA (siRNA) or P2RX7 siRNA (80 pmol/mL), followed by exposure to LL-37. TUBB3 mRNA expression. (C) CAMP, (D) FPRL1, and (E) P2RX7 mRNA expression in human colon cancer PCR array plate. (F) Cell migration of SW620 cells. SW620 cells were treated with DMSO, KN62, and WRW4 for 30 min, followed by LL-37 for 7 h. Results were pooled from three independent experiments.

Article Snippet: Human P2RX7 , Hs00175721_m1.

Techniques: Inhibition, Migration, Transfection, Control, Small Interfering RNA, Expressing

Journal: Molecular Therapy Oncolytics

Article Title: Cathelicidin Suppresses Colon Cancer Metastasis via a P2RX7-Dependent Mechanism

doi: 10.1016/j.omto.2019.01.004

Figure Lengend Snippet: Cathelicidin Inhibited TUBB3 Expression via P2RX7 (A) Green tubulin tracker staining with blue nuclear staining in human cancer SW620 cells. SW620 cells were pretreated with DMSO (10 μL/mL) or KN62 (10 μM) for 30 min, followed by exposure to LL-37 (5 μM) for 24 h. LL-37 reduced tubulin expression in SW620 cells that was prevented by KN62. (B) SW620 cells were transiently transfected with control small interfering RNA (siRNA) or P2RX7 shRNA (1 μg/mL), followed by exposure to LL-37. TUBB3 mRNA expression in SW620 cells. (C) P2RX7 mRNA expression in the transfected SW620 cells. (D) HT-29 cells were pretreated with DMSO (10 μL/mL) or KN62 (10 μM) for 30 min, followed by exposure to LL-37. TUBB3 mRNA expression. Results were pooled from three independent experiments.

Article Snippet: Human P2RX7 , Hs00175721_m1.

Techniques: Expressing, Staining, Transfection, Control, Small Interfering RNA, shRNA

Journal: Molecular Therapy Oncolytics

Article Title: Cathelicidin Suppresses Colon Cancer Metastasis via a P2RX7-Dependent Mechanism

doi: 10.1016/j.omto.2019.01.004

Figure Lengend Snippet: Cathelicidin Inhibited Colon Cancer Metastasis via P2RX7 (A and B) Human cytokeratin-18 expression (representing human colon cancer cells) in (A) lungs and (B) liver of nude mice was identified by brown color spots (indicated by arrows). Intravenous cathelicidin expressing AAV administration reduced human CK18 expression in lungs and liver of nude mice.

Article Snippet: Human P2RX7 , Hs00175721_m1.

Techniques: Expressing

Journal: Molecular Therapy Oncolytics

Article Title: Cathelicidin Suppresses Colon Cancer Metastasis via a P2RX7-Dependent Mechanism

doi: 10.1016/j.omto.2019.01.004

Figure Lengend Snippet: Cathelicidin Inhibited TUBB3 mRNA Expression in Metastasized Tumors via P2RX7 (A and B) Human keratin 20 mRNA expression (human colon cancer cell marker) in (A) lungs and (B) liver of nude mice was significantly reduced by CAMP -HA-AAV. Intraperitoneal KN62 treatment increased the presence of cytokeratin 18 protein and keratin 20 mRNA expression in the CAMP -HA-AAV-treated nude mice. (C and D) Human TUBB3 mRNA expression in (C) lungs and (D) liver of nude mice was significantly reduced by CAMP -HA-AAV. Intraperitoneal KN62 treatment increased the presence of human TUBB3 mRNA expression in the CAMP -HA-AAV-treated nude mice. (E) Diameters of subcutaneous tumors. Intravenous CAMP -HA-AAV significantly reduced subcutaneous tumor diameters in nude mice that were partially reversed by KN62 treatment.

Article Snippet: Human P2RX7 , Hs00175721_m1.

Techniques: Expressing, Marker

Journal: Molecular Therapy Oncolytics

Article Title: Cathelicidin Suppresses Colon Cancer Metastasis via a P2RX7-Dependent Mechanism

doi: 10.1016/j.omto.2019.01.004

Figure Lengend Snippet: Primers for Real-Time RT-PCR

Article Snippet: Human P2RX7 , Hs00175721_m1.

Techniques:

Journal: Journal of cell science

Article Title: Autocrine regulation of TGF-β1-induced cell migration by exocytosis of ATP and activation of P2 receptors in human lung cancer cells.

doi: 10.1242/jcs.104976

Figure Lengend Snippet: Fig. 4. Involvement of P2X7 receptor in TGF-b1-induced migration of A549 cells. (A) A549 cells were transfected with 10 nM P2X7 siRNA or control siRNA and incubated for 60 hours. Then the expression of P2X7 receptor was detected by immunoblotting as described in Materials and Methods. (B,C) 60 hours after transfection, TGF-b1-induced cell migration was examined using Culture-Insert (B) and Transwell (C) systems, as described in Materials and Methods. Values are means 6 s.e.m. (n56). **P,0.01: significant differences between the test groups and control group. (D) 60 hours after transfection, cells were incubated with or without TGF-b1 (5 ng/ml) for 12 hours. Then F-actin was stained using Rhodamine– phalloidin (red) and stained cells were analyzed using a confocal laser- scanning microscope at 636 magnification. To verify the location of nuclei, cells were stained with Hoechst33258 (blue).

Article Snippet: A549 cells were transfected with 10 nM P2X7 siRNA (a pool of three targetspecific 19–25 nt siRNAs) or control siRNA-A (Santa Cruz) using HiPerFect Transfection Reagent (Qiagen, Valencia, CA) according to the manufacturers’ instructions.

Techniques: Migration, Transfection, Control, Incubation, Expressing, Western Blot, Staining, Laser-Scanning Microscopy

Journal: Journal of cell science

Article Title: Autocrine regulation of TGF-β1-induced cell migration by exocytosis of ATP and activation of P2 receptors in human lung cancer cells.

doi: 10.1242/jcs.104976

Figure Lengend Snippet: Fig. 7. Decrease in ATP-enriched vesicles in A549 cells by knockdown of SLC17A9. (A) A549 cells were transfected with 2 mg shRNA targeting SLC17A9 (red or green) or scrambled shRNA. Then 48 hours after transfection, total RNA was extracted and SLC17A9 gene expression was examined by measuring mRNA levels with real-time RT-PCR. (B) Cell membrane proteins were extracted 72 hours after transfection, and the expression of SLC17A9 was detected by immunoblotting as described in Materials and Methods. SLC17A9 expression levels were normalized to EGFR expression levels and expressed relative to those of scrambled shRNA-transfected cells. (C) 72 hours after transfection, expression of SLC17A9 protein was detected by immunocytochemistry. (D) ATP-enriched vesicles were detected using MANT-ATP (cyan, upper panels) and quinacrine dihydrochloride (magenta, middle panels) 72 hours after transfection. The merged images are shown in lower panels.

Article Snippet: A549 cells were transfected with 10 nM P2X7 siRNA (a pool of three targetspecific 19–25 nt siRNAs) or control siRNA-A (Santa Cruz) using HiPerFect Transfection Reagent (Qiagen, Valencia, CA) according to the manufacturers’ instructions.

Techniques: Knockdown, Transfection, shRNA, Gene Expression, Quantitative RT-PCR, Membrane, Expressing, Western Blot, Immunocytochemistry

Journal: Journal of cell science

Article Title: Autocrine regulation of TGF-β1-induced cell migration by exocytosis of ATP and activation of P2 receptors in human lung cancer cells.

doi: 10.1242/jcs.104976

Figure Lengend Snippet: Fig. 8. Involvement of SLC17A9-dependent ATP release in TGF-b1-induced migration and actin remodeling. (A) Cells were transfected with shRNA targeting SLC17A9 (red or green) or scrambled shRNA and incubated for 72 hours. The transfected cells were stimulated with TGF-b1 (5 ng/ml) and further incubated for the indicated times; then the concentration of ATP in the culture medium was measured as described in Materials and Methods. (B,C) 72 hours after transfection, TGF-b1-induced cell migration was examined using Culture-Insert (B) and Transwell (C) systems, as described in Materials and Methods. The transfected cells were incubated with vehicle or TGF-b1 (5 ng/ml) for another 24 hours. Values are means 6 s.e.m. (n58). **P,0.01: significant differences between the test groups and control group. (D) 72 hours after transfection, cells were incubated with or without TGF-b1 (5 ng/ml) for 12 h. Then F-actin was stained using Rhodamine–phalloidin (red) and stained cells were analyzed using a confocal laser-scanning microscope at 636 magnification. To verify the location of nuclei, cells were stained with Hoechst33258 (blue). (E) Non-transfected A549 cells were incubated with TGF-b1 (5 ng/ml) for the indicated times. Then, cell membrane proteins were extracted and expression of E-cadherin and N-cadherin was detected by immunoblotting as described in Materials and Methods. (F) 72 hours after transfection, cells were incubated with or without TGF-b1 for another 48 hours and expression of E-cadherin and N-cadherin was detected. EGFR was detected as a loading control.

Article Snippet: A549 cells were transfected with 10 nM P2X7 siRNA (a pool of three targetspecific 19–25 nt siRNAs) or control siRNA-A (Santa Cruz) using HiPerFect Transfection Reagent (Qiagen, Valencia, CA) according to the manufacturers’ instructions.

Techniques: Migration, Transfection, shRNA, Incubation, Concentration Assay, Control, Staining, Laser-Scanning Microscopy, Membrane, Expressing, Western Blot

Journal: Journal of neuroinflammation

Article Title: Methamphetamine alters microglial immune function through P2X7R signaling.

doi: 10.1186/s12974-016-0553-3

Figure Lengend Snippet: Fig. 1 METH increases P2X7 purinergic receptor mRNA and protein expression in microglia. a The expression level of P2X7R mRNA in ESdM cells was modestly increased by treatment with 100 μM METH for 48 h (P < 0.0336). b Representative immunoblots of P2X7R in whole cell lysate, protein fraction, and Ponceau S staining (Po-S) as loading control. The ratios of P2X7R to protein loading control are shown in the histogram. Error bars represent mean ± SE of four independent experiments (p < 0.0003). (METH-treated versus control)

Article Snippet: The images were analyzed with AxioVision version 4.7 software (Carl Zeiss MicroImaging) and with National Institutes of Health ImageJ version 1.42 software (http://rsbweb.nih.gov/ij/) as described [25]. siRNA knockdown of P2X7R P2X7R expression in ESdM cells was silenced by small interfering RNA (siRNA) transfection for about 48 h. P2rx7 Trilencer-27 Mouse siRNA (OriGene Technologies; Rockville, MD) was used in conjunction with the jetPRIME transfection reagent (Polypus transfectionTM; Bioparc, France) to directly transfect the siRNA in N2 medium according to the transfection protocol.

Techniques: Expressing, Western Blot, Staining, Control

Journal: Journal of neuroinflammation

Article Title: Methamphetamine alters microglial immune function through P2X7R signaling.

doi: 10.1186/s12974-016-0553-3

Figure Lengend Snippet: Fig. 3 METH increases phagocytic capacity of microglia that is P2X7-dependent. Graphical representation of microglial phagocytosis in response to treatment with Cyto D (5 μM), A 438079 (10 μM), METH (100 μM), or METH (100 μM), and A 438079 (10 μM) was quantified using flow cytometry. METH (100 μM) significantly increases microglia phagocytosis (****p < 0.001). Pretreatment with antagonist decreases phagocytosis as compared to METH-treated cells. (****p < 0.0001) (ANOVA). Data consist of means ± SEM of three independent experiments

Article Snippet: The images were analyzed with AxioVision version 4.7 software (Carl Zeiss MicroImaging) and with National Institutes of Health ImageJ version 1.42 software (http://rsbweb.nih.gov/ij/) as described [25]. siRNA knockdown of P2X7R P2X7R expression in ESdM cells was silenced by small interfering RNA (siRNA) transfection for about 48 h. P2rx7 Trilencer-27 Mouse siRNA (OriGene Technologies; Rockville, MD) was used in conjunction with the jetPRIME transfection reagent (Polypus transfectionTM; Bioparc, France) to directly transfect the siRNA in N2 medium according to the transfection protocol.

Techniques: Flow Cytometry

Journal: PloS one

Article Title: P2X4 assembles with P2X7 and pannexin-1 in gingival epithelial cells and modulates ATP-induced reactive oxygen species production and inflammasome activation.

doi: 10.1371/journal.pone.0070210

Figure Lengend Snippet: Figure 3. Involvement of P2X4, P2X7, and pannexin-1 in ATP-mediated ROS generation. (A) Total RNA from immortalized GEC was extracted and reversed transcribed (RT) to cDNA. The cDNA was used to perform PCR with the primers specific for the indicated genes, and the PCR products were finally visualized by EtBr staining. (B) GEC were treated with 100 mM or 3 mM ATP as indicated for 1 hour, and ROS production was measured with DCF staining and visualized by fluorescence microcopy. Quantification of the fluorescence in three independent experiments with SD is shown in (C). (D–E) ROS production was measured by DCF staining in GEC stimulated under different conditions. GEC were left untreated or stimulated with 20 mM nigericin or 3 mM ATP for 1 hour followed by fluorescence microscopy. Diminished ROS generation by different receptor antagonists was examined by pretreating cells with 100 mM oxATP for 30 minutes, 1 mM probenecid for 10 minutes, or 50 mM 5-BDBD or 100 mM PPADS for 15 minutes, followed by 3 mM ATP stimulation for 1 hour. doi:10.1371/journal.pone.0070210.g003

Article Snippet: The siRNA sequences were: 59-GCUUUCAACGGGUCUGUCATT39 and 59-UGACAGACCCGUUGAAAGCTA-39 for P2X4 (Ambion, LifeCell Technologies, S9957, Cat. #: 4392420); and 59-ACAAUGUUGAGAAACGGACUCUGAT-39 for P2X7 (27 mer siRNA duplexes OriGene Technologies, Cat. #: SR303325).

Techniques: Staining, Fluorescence, Microscopy

Journal: PloS one

Article Title: P2X4 assembles with P2X7 and pannexin-1 in gingival epithelial cells and modulates ATP-induced reactive oxygen species production and inflammasome activation.

doi: 10.1371/journal.pone.0070210

Figure Lengend Snippet: Figure 4. Diminished ATP-induced ROS production due to depletion of P2X4 or P2X7 by RNA interference. (A) Immortalized GEC were transduced with lentiviruses carrying the indicated shRNA-expressing plasmid for 1 day and selected with puromycin. After selection, cells were collected and total RNA was analyzed by qPCR to confirm knockdown efficiency. (B) DCF staining of ROS production after 3 mM ATP stimulation for 1 hour in different cell lines. The fluorescence shown in (C) was quantified as in Figure 1 and normalized to shCtrl, which was transduced with control, non-mammalian shRNA. doi:10.1371/journal.pone.0070210.g004

Article Snippet: The siRNA sequences were: 59-GCUUUCAACGGGUCUGUCATT39 and 59-UGACAGACCCGUUGAAAGCTA-39 for P2X4 (Ambion, LifeCell Technologies, S9957, Cat. #: 4392420); and 59-ACAAUGUUGAGAAACGGACUCUGAT-39 for P2X7 (27 mer siRNA duplexes OriGene Technologies, Cat. #: SR303325).

Techniques: Transduction, shRNA, Expressing, Plasmid Preparation, Selection, Knockdown, Staining, Fluorescence, Control

Journal: PloS one

Article Title: P2X4 assembles with P2X7 and pannexin-1 in gingival epithelial cells and modulates ATP-induced reactive oxygen species production and inflammasome activation.

doi: 10.1371/journal.pone.0070210

Figure Lengend Snippet: Figure 5. Impaired ATP-stimulated caspase-1 activation due to disruption of the complex containing P2X4, P2X7, and pan- nexin-1. (A) Immortalized GEC that had been depleted of P2X4 or P2X7 using lentiviral particles were treated with 100 mM or 3 mM ATP for 3 hours, and supernatants were collected for caspase-1 activity measure- ment. Caspase-1 activity was measured by ELISA as described in Material and Methods. (B) Total proteins isolated from GEC were subjected to immunoprecipitation (IP) by a polyclonal anti-P2X4 antibody or Dynabeads as a control. Precipitates or total protein extract (as input) were resolved on SDS-PAGE and analyzed on immunoblots with anti-P2X7 (top), anti-pannexin-1 (middle), or anti- P2X4 (bottom) antibodies. doi:10.1371/journal.pone.0070210.g005

Article Snippet: The siRNA sequences were: 59-GCUUUCAACGGGUCUGUCATT39 and 59-UGACAGACCCGUUGAAAGCTA-39 for P2X4 (Ambion, LifeCell Technologies, S9957, Cat. #: 4392420); and 59-ACAAUGUUGAGAAACGGACUCUGAT-39 for P2X7 (27 mer siRNA duplexes OriGene Technologies, Cat. #: SR303325).

Techniques: Activation Assay, Disruption, Activity Assay, Enzyme-linked Immunosorbent Assay, Isolation, Immunoprecipitation, Control, SDS Page, Western Blot

Journal: PloS one

Article Title: P2X4 assembles with P2X7 and pannexin-1 in gingival epithelial cells and modulates ATP-induced reactive oxygen species production and inflammasome activation.

doi: 10.1371/journal.pone.0070210

Figure Lengend Snippet: Figure 6. Abrogation of ATP-induced IL-1b secretion in P. gingivalis-infected GEC by inhibition of P2X4, P2X7, or pannexin-1. Primary GEC (C and D) and immortalized GEC (A and B) were infected with or without P. gingivalis (P.g.) at an M.O.I. of 100 for 6 hours, followed by treatment with different pharmaceutical agents. Infected cells were treated with 100 mM ATP, 3 mM ATP, 3 mM ADP, 3 mM AMP, or 3 mM UTP individually for 1

Article Snippet: The siRNA sequences were: 59-GCUUUCAACGGGUCUGUCATT39 and 59-UGACAGACCCGUUGAAAGCTA-39 for P2X4 (Ambion, LifeCell Technologies, S9957, Cat. #: 4392420); and 59-ACAAUGUUGAGAAACGGACUCUGAT-39 for P2X7 (27 mer siRNA duplexes OriGene Technologies, Cat. #: SR303325).

Techniques: Infection, Inhibition

Journal: PloS one

Article Title: P2X4 assembles with P2X7 and pannexin-1 in gingival epithelial cells and modulates ATP-induced reactive oxygen species production and inflammasome activation.

doi: 10.1371/journal.pone.0070210

Figure Lengend Snippet: Figure 7. Model showing the role of P2X4 and P2X7 in ROS production and inflammasome activation in GEC stimulated with extracellular ATP. doi:10.1371/journal.pone.0070210.g007

Article Snippet: The siRNA sequences were: 59-GCUUUCAACGGGUCUGUCATT39 and 59-UGACAGACCCGUUGAAAGCTA-39 for P2X4 (Ambion, LifeCell Technologies, S9957, Cat. #: 4392420); and 59-ACAAUGUUGAGAAACGGACUCUGAT-39 for P2X7 (27 mer siRNA duplexes OriGene Technologies, Cat. #: SR303325).

Techniques: Activation Assay