Journal: Cells

Article Title: Upregulation of TLR4-Dependent ATP Production Is Critical for Glaesserella parasuis LPS-Mediated Inflammation

doi: 10.3390/cells12050751

Figure Lengend Snippet: ATP induces inflammation and increases P2X7 expression. ( A , B ) ELISA analysis of IL-1β normalized to the control. PAM cells were treated with G. parasuis LZ ( A ) /50 μg/mL LPS ( B ) in the presence and absence of ATP, apyrase, and nigericin. ( C ) mRNA expression measured by qRT-PCR for IL-1β level normalized to the control. ( D ) Western blot analysis of P2X7R, NLRP3, NF-κB, p-NF-κB, and GSDMD expression in PAM cells. Cells were treated with or without LPS in the presence and absence of nigericin. All proteins were normalized to the level of β-actin. Data represent mean ± SEM, n = 3, * p < 0.05, ** p < 0.01.

Article Snippet: The main antibodies were mouse anti-β-actin antibodies and those against NF-κB, p-NF-κB, GSDMD, NLRP3, IL-1β, caspase1, and P2X7 receptor from Cell Signaling Technology in the United States.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control, Quantitative RT-PCR, Western Blot

Journal: Cells

Article Title: Upregulation of TLR4-Dependent ATP Production Is Critical for Glaesserella parasuis LPS-Mediated Inflammation

doi: 10.3390/cells12050751

Figure Lengend Snippet: A740003 regulates P2X7 function and inhibits inflammation. ( A ) Western blot analysis of P2X7R, NLRP3 expression in PAM cells. All proteins were normalized to the level of β-actin. Cells were treated with or without 50 μg/mL LPS in the presence and absence of A-740003 (10 μM) ( B ) Western blot analysis of NF-κB, p- NF-κB, and GSDMD expression in PAM cells. All proteins were normalized to the level of β-actin. ( C ) ELISA analysis of IL-1β normalized to the control. ( D ) Quantification of mortality. After PAM cells were treated, cell viability was measured by CCK-8. ( E ) Representative images of immunofluorescence staining. Differentiated PAM cells were treated with or without LPS in the presence and absence of 0.1 μM A-740003. Nuclei were stained by DAPI in Blue and NF-kB p65 were stained in green, then observed using an inverted fluorescence microscope, 100×. Data represent mean ± SEM, n = 3, * p < 0.05, ** p < 0.01.

Article Snippet: The main antibodies were mouse anti-β-actin antibodies and those against NF-κB, p-NF-κB, GSDMD, NLRP3, IL-1β, caspase1, and P2X7 receptor from Cell Signaling Technology in the United States.

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Control, CCK-8 Assay, Immunofluorescence, Staining, Fluorescence, Microscopy

Journal: Biology Open

Article Title: P2X 7 regulates ependymo-radial glial cell proliferation in adult Danio rerio following spinal cord injury

doi: 10.1242/bio.060270

Figure Lengend Snippet: P2X 7 receptors were expressed by GFAP + ependymo-radial glial cells. Spinal cord cross sections are shown (dorsal is up; asterisk indicates central canal; scale bar: 50 μm). Representative images of GFAP and P2X 7 labelling in naïve fish (A-C′), at 7 dpi (D-F′), and at 14 dpi (G-I′). Colocalization (arrows) was seen in radial processes and cell bodies (C′,F′,I′). GFAP + cells that did not colocalize with P2X 7 were also detected (open arrowheads) (C′,F′,I′). n =3/group. Created with BioRender.com.

Article Snippet: This included a P2X 7 control antigen (Alomone Labs, BLP-PR004, Q64663).

Techniques:

Journal: Biology Open

Article Title: P2X 7 regulates ependymo-radial glial cell proliferation in adult Danio rerio following spinal cord injury

doi: 10.1242/bio.060270

Figure Lengend Snippet: P2X 7 receptors were expressed by PCNA + proliferating cells. Spinal cord cross sections are shown (dorsal is up; asterisk indicates central canal; scale bar: 50 μm). Representative images of PCNA + and P2X 7 labelling in naïve fish (A-C′), at 7 dpi rostral (D-F′), and at 7 dpi caudal (G-I′). Colocalization (arrows) was seen in some cells (C′,F′,I′). PCNA + cells that did not colocalize with P2X 7 were also detected (open arrowheads) (C′,F′,I′). n =3/group. Created with BioRender.com.

Article Snippet: This included a P2X 7 control antigen (Alomone Labs, BLP-PR004, Q64663).

Techniques:

Journal: Biology Open

Article Title: P2X 7 regulates ependymo-radial glial cell proliferation in adult Danio rerio following spinal cord injury

doi: 10.1242/bio.060270

Figure Lengend Snippet: P2X 7 receptors were expressed by HuC/D + neurons. Spinal cord cross sections are shown (dorsal is up; asterisk indicates central canal; scale bar: 50 μm). Representative images of HuC/D + and P2X 7 labelling in naïve fish (A-C′), at 14 dpi rostral (D-F′), and at 14 dpi caudal (G-I′). Colocalization (arrows) was seen in larger mature neurons and smaller immature neurons (C′,F′,I′). n =3/group. Created with BioRender.com.

Article Snippet: This included a P2X 7 control antigen (Alomone Labs, BLP-PR004, Q64663).

Techniques:

Journal: Biology Open

Article Title: P2X 7 regulates ependymo-radial glial cell proliferation in adult Danio rerio following spinal cord injury

doi: 10.1242/bio.060270

Figure Lengend Snippet: Temporal changes in P2X 7 protein expression following SCI in adult zebrafish. Schematic representations of naïve and injured spinal cord areas of analysis (A-B), as well as tissue collection timeline (C). Representative western blot and corresponding total protein in naïve zebrafish (Zf) spinal cord tissue, mouse (Ms) hippocampal tissue as a positive control (+), and pre-absorption of naïve zebrafish spinal cord tissue with a blocking peptide as a negative control (-) (D). Representative western blot and corresponding total protein at 1 dpi (E), 7 dpi (G), and 14 dpi (I). Quantitative analysis of the 50 kDa isoform showed significant downregulation within the injury (In, n =10, one-way ANOVA with Dunnett's post hoc test, P =0.0079) and caudal to the lesion (Ca, n =9, P =0.0046) at 7 dpi when normalized to naïve tissue ( n =9) (H). Protein expression of the 50 kDa isoform showed basal levels of expression at 1 dpi (F) and 14 dpi (J). Data presented as means±s.d. **, significant differences, P <0.01. Created with BioRender.com.

Article Snippet: This included a P2X 7 control antigen (Alomone Labs, BLP-PR004, Q64663).

Techniques: Expressing, Western Blot, Positive Control, Blocking Assay, Negative Control

Journal: Journal of Inflammation (London, England)

Article Title: Effects of long non-coding RNA Gm14461 on pain transmission in trigeminal neuralgia

doi: 10.1186/s12950-019-0231-1

Figure Lengend Snippet: Effect of Gm14461 expression on MWT and expression of Gm14461, CGRP, P2X3 receptor, P2X7 receptor, TNF-α, IL-1β, and IL-6 in TN mice. C57BL/6 J mice were randomly divided into six groups ( n = 8/group): Sham, TN, TN + Scramble siRNA, TN + si-Gm14461, TN + Vector, and TN + Gm14461 group. a Gm14461 expression in mouse TGs was detected by qRT-PCR. b The MWT was measured to assess the nociception of mice at different times after CCI-ION surgery (0, 1, 3, 5, 7, 9, 11, 13, 15 d). * P < 0.05: Sham vs. TN; # P < 0.05 Scramble siRNA vs. si-Gm14461; $ P < 0.05 Vector vs. Gm14461. c The protein levels of CGRP, P2X3 receptor, and P2X7 receptor were examined using western blot. d-f The mRNA levels of TNF-α ( d ), IL-1β (e), and IL-6 (f) in mouse TGs were detected by qRT-PCR. Data are presented as mean ± SD. * P < 0.05 vs. Sham; # P < 0.05 vs. Scramble siRNA; $ P < 0.05 vs. Vector

Article Snippet: The membrane was then blocked with 5% non-fat dry milk and incubated with the primary antibodies against the CGRP, P2X3, and P2X7 (1:1000; all purchased from Santa Cruz Biotechnology).

Techniques: Expressing, Plasmid Preparation, Quantitative RT-PCR, Western Blot

Journal: Journal of Inflammation (London, England)

Article Title: Effects of long non-coding RNA Gm14461 on pain transmission in trigeminal neuralgia

doi: 10.1186/s12950-019-0231-1

Figure Lengend Snippet: Effect of Gm14461 expression on the expression of CGRP, P2X3 receptor, and P2X7 receptor in primary TGNs. a The primary mouse TGNs were isolated from C57BL/6 J mice and treated with TNF-α (10 ng/mL), IL-1β (25 ng/mL) and IL-6 (25 ng/mL) for 24 h. Gm14461 expression in TGNs was examined by qRT-PCR. * P < 0.05 vs. Control. b Western blot analysis of protein levels of CGRP, P2X3 receptor, and P2X7 receptor in primary TGNs transfected with scramble siRNA, si-Gm14461, empty vector, and pcDNA3.1-Gm14461 in the stimulation of TNF-α (10 ng/mL) for 24 h. c The qualifications of western blots in ( b ). Data are presented as mean ± SD. N = 3. * P < 0.05 vs. Control; # P < 0.05 vs. TNF-α + Scramble siRNA; $ P < 0.05 vs. TNF-α + Vector

Article Snippet: The membrane was then blocked with 5% non-fat dry milk and incubated with the primary antibodies against the CGRP, P2X3, and P2X7 (1:1000; all purchased from Santa Cruz Biotechnology).

Techniques: Expressing, Isolation, Quantitative RT-PCR, Control, Western Blot, Transfection, Plasmid Preparation

Journal: Purinergic Signalling

Article Title: P2X7 receptor: the regulator of glioma tumor development and survival

doi: 10.1007/s11302-021-09834-2

Figure Lengend Snippet: Characterization of P2X7 receptor in glioma C6 cells in vitro. a Western blot analysis of P2X7 protein expression in C6 cells after P2X7 silencing for 72 h ( n = 3). b Calcium signal level in glioma C6 cells after P2X7 downregulation and BzATP stimulation (300 µM) ( n = 4). Gray lines represent the signals from the biological replicates; black lines represent the signal averaged among the individual replicates. Insert shows effect of P2X7 downregulation on the signal strength, one-way t -test: * P ≤ 0.05. c Measurement of bioluminescence originated from released ATP after P2X7 activation/inhibition in C6 cells ( n = 3). The statistical significance of the differences was determined with one-way ANOVA with Bonferroni post hoc test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. control group

Article Snippet: Three target-specific RNAi (small interfering RNA) complementary to rat P2X7 transcript (sc-108056) and control non-silencing RNAi were obtained from Santa Cruz Biotechnology, Inc. (USA).

Techniques: In Vitro, Western Blot, Expressing, Activation Assay, Inhibition, Control

Journal: Purinergic Signalling

Article Title: P2X7 receptor: the regulator of glioma tumor development and survival

doi: 10.1007/s11302-021-09834-2

Figure Lengend Snippet: P2X7 activation increased C6 cell proliferation, viability, and cell adhesion in vitro. Quantitative data are presented as signal relative to control. a MTS test of long-term C6 cell viability after 100 µM BzATP stimulation of control cells and cells with P2X7 downregulation using RNAi ( n = 3). The significance of the differences was determined with paired t -test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. the respective control. b Representative Western blot analysis of pro-survival and pro-proliferative proteins: p-p38 (Thr180/Tyr182), pAkt (Ser473), tAkt, CD133, HSPA1, and HSPA5 in C6 cell lysates after 24-h BzATP stimulation ( n = 3). c Upper panel: MTS test of C6 cell viability after 200 µM BCNU (carmustine) treatment for 24 h. Co-treatment with 100 µM BzATP and 200 µM BCNU increased viability of C6 cells ( n = 3). The significance of the differences was determined with one-way ANOVA with Bonferroni post hoc test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. all groups. Bottom panel: a crystal violet assay demonstrating a synergistic effect of BBG with BCNU/TMZ co-treatment on C6 cell growth for 24 h ( n = 6). The significance of the differences was determined with repeated measures one-way ANOVA with Duncan’s multiple range post hoc test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. all groups. d P2X7 activation affects C6 cell adhesion to extracellular matrix components: collagen I, collagen IV, fibronectin. C6 cells showed higher adhesion potential after 24-h BzATP stimulation. 24-h co-treatment/treatment with 100 nM BBG significantly decreased C6 cell adhesion to ECM components ( n = 6). The significance of the differences was determined with one-way ANOVA with Bonferroni post hoc test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. control group. e P2X7 RNAi downregulation declined C6 cell adhesion to ECM components upon BzATP stimulation ( n = 4). The significance of the differences was determined with paired t -test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. the respective control

Article Snippet: Three target-specific RNAi (small interfering RNA) complementary to rat P2X7 transcript (sc-108056) and control non-silencing RNAi were obtained from Santa Cruz Biotechnology, Inc. (USA).

Techniques: Activation Assay, In Vitro, Control, Western Blot, Crystal Violet Assay

Journal: Purinergic Signalling

Article Title: P2X7 receptor: the regulator of glioma tumor development and survival

doi: 10.1007/s11302-021-09834-2

Figure Lengend Snippet: Activation of P2X7 receptor led to elevated ROS production and mitochondrial membrane depolarization in C6 cells. Quantitative data are presented as signal relative to control. a DCF-DA fluorescence measurement in C6 glioma cells. Cells stimulated with 100 µM BzATP for 1 h were characterized by increased ROS production compared to control in C6 cells ( n = 3). The significance of the differences was determined with one-way ANOVA with Bonferroni post hoc test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. control group. b Potential-dependent staining of mitochondria in C6 cells using JC-1. 1-h BzATP (100 µM) stimulation led to considerable mitochondrial membrane depolarization ( n = 3). The significance of the differences was determined with one-way ANOVA with Bonferroni post hoc test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. control group. c Representative images of MitoTracker Red CMXRos accumulation in C6 cells stimulated with 100 µM BzATP for 1 h. d DCF-DA fluorescence measurement in C6 glioma cells with RNA interference of P2X7 expression ( n = 4). Downregulation of P2X7 led to decreased ROS production in C6 cells after 1-h BzATP (100 µM) stimulation. The significance of the differences was determined using paired t -test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. the respective control. e Potential-dependent staining of mitochondria using JC-1 dye in C6 cells with RNA interference of P2X7 expression after 1 h of 100 μM BzATP pre-treatment ( n = 3). The amount of depolarized mitochondria was higher in control cells when compared to that in cells with P2X7 downregulation. The significance of the differences was determined using paired t -test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. the respective control

Article Snippet: Three target-specific RNAi (small interfering RNA) complementary to rat P2X7 transcript (sc-108056) and control non-silencing RNAi were obtained from Santa Cruz Biotechnology, Inc. (USA).

Techniques: Activation Assay, Membrane, Control, Fluorescence, Staining, Expressing

Journal: Purinergic Signalling

Article Title: P2X7 receptor: the regulator of glioma tumor development and survival

doi: 10.1007/s11302-021-09834-2

Figure Lengend Snippet: Effect of P2X7 inhibition using BBG on glioma C6 tumor development in vivo. a Brilliant Blue G administration led to reduction of glioma tumor mass. Representative images of control and BBG-treated C6 glioma tumors ( n = 9 for control group and n = 10 for BBG-treated group). The significance of the differences was determined using Student’s t -test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. the respective control. b Representative Western blot showing the significant decrease of P2X7 expression in tumor homogenates after BBG treatment. Representative image of P2X7 immunodetection in C6 glioma tumors. c P2X7 inhibition resulted in diminished activation of matrix metalloproteinase-2 in glioma tumors. The significance of the differences was determined with Student’s t -test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. the respective control. d Western blot analysis of known proteins involved in cell adhesion and EMT signaling in C6 tumor homogenates ( n = 4). e Western blot analysis of known proteins involved in tumor aggressiveness in C6 tumor homogenates ( n = 4)

Article Snippet: Three target-specific RNAi (small interfering RNA) complementary to rat P2X7 transcript (sc-108056) and control non-silencing RNAi were obtained from Santa Cruz Biotechnology, Inc. (USA).

Techniques: Inhibition, In Vivo, Control, Western Blot, Expressing, Immunodetection, Activation Assay

Journal: Purinergic Signalling

Article Title: P2X7 receptor: the regulator of glioma tumor development and survival

doi: 10.1007/s11302-021-09834-2

Figure Lengend Snippet: P2X7 inhibition reduced the ATP release with simultaneous decrease of some pro-inflammatory cytokine levels in mouse serum and changed the redox metabolism in C6 tumors. a An ATP release is significantly lower in tumors treated with BBG compared to the control tumors ( n = 4). The significance of the differences was determined using Student’s t -test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. the respective control. b Decline NOS-2 expression in BBG-treated tumors ( n = 4). c Decreased expression of FOXP3 (T reg lymphocytes) and CD68 (macrophages) markers in BBG-treated tumors evaluated using flow cytometry ( n = 5). The significance of the differences was determined using Student’s t -test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. the respective control. d The level of the pro-inflammatory and anti-inflammatory cytokines, in mouse serum and glioma tumors using cytokine array for multiplex protein detection ( n = 3). e P2X7 inhibition caused a decrease of GSSG/GSH ratio with a slight decline of SOD activity in BBG-treated tumors ( n = 4). The significance of the differences was determined using Student’s t -test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. the respective control

Article Snippet: Three target-specific RNAi (small interfering RNA) complementary to rat P2X7 transcript (sc-108056) and control non-silencing RNAi were obtained from Santa Cruz Biotechnology, Inc. (USA).

Techniques: Inhibition, Control, Expressing, Flow Cytometry, Multiplex Assay, Activity Assay

Journal: Purinergic Signalling

Article Title: P2X7 receptor: the regulator of glioma tumor development and survival

doi: 10.1007/s11302-021-09834-2

Figure Lengend Snippet: P2X7 receptor effects on human glioma development. a Western blot analysis of p-p38 MAPK, CD-133 and HSPA1 expression in U-138 human glioma cells ( n = 3). b In silico analysis of P2X7 expression level and survival of glioma patients using information from the TCGA database. c In silico analysis of P2X7 expression level in different WHO astrocytoma grades compared to normal tissue using NCBI dataset. d A transwell migration assay of LN-229 and U-251 glioma cells treated with 2 μM KN-62 for 24 h ( n = 3). e A crystal violet assay demonstrating a synergistic effect of 2 μM KN-62 with BCNU/TMZ co-treatment on LN-229, U-251, and U-138 glioma cell growth for 24 h ( n = 4). The significance of the differences was determined with repeated-measures one-way ANOVA with Duncan multiple range post hoc test: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 vs. all groups

Article Snippet: Three target-specific RNAi (small interfering RNA) complementary to rat P2X7 transcript (sc-108056) and control non-silencing RNAi were obtained from Santa Cruz Biotechnology, Inc. (USA).

Techniques: Western Blot, Expressing, In Silico, Transwell Migration Assay, Crystal Violet Assay

Journal: PLoS ONE

Article Title: P2X 4 Assembles with P2X 7 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: (A) Immortalized GEC that had been depleted of P2X 4 or P2X 7 using lentiviral particles were treated with 100 µM or 3 mM ATP for 3 hours, and supernatants were collected for caspase-1 activity measurement. Caspase-1 activity was measured by ELISA as described in . (B) Total proteins isolated from GEC were subjected to immunoprecipitation (IP) by a polyclonal anti-P2X 4 antibody or Dynabeads as a control. Precipitates or total protein extract (as input) were resolved on SDS-PAGE and analyzed on immunoblots with anti-P2X 7 (top), anti-pannexin-1 (middle), or anti-P2X 4 (bottom) antibodies.

Article Snippet: The siRNA sequences were: 5′-GCUUUCAACGGGUCUGUCATT-3′ and 5′-UGACAGACCCGUUGAAAGCTA-3′ for P2X 4 (Ambion, LifeCell Technologies, S9957, Cat. #: 4392420); and 5′-ACAAUGUUGAGAAACGGACUCUGAT-3′ for P2X 7 (27 mer siRNA duplexes OriGene Technologies, Cat. #: SR303325).

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

Journal: PLoS ONE

Article Title: P2X 4 Assembles with P2X 7 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: 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 µM ATP, 3 mM ATP, 3 mM ADP, 3 mM AMP, or 3 mM UTP individually for 1 hour (A and C). Alternatively, infected cells were pre-treated with 50 µM 5-BDBD for 15 minutes, 100 µM PPADS for 15 minutes, 100 µM oxATP for 30 minutes, or 1 mM probenecid for 10 minutes, followed by treatment with 3 mM ATP for 1 hour (B and D). The supernatants were collected and subjected to ELISA to measure IL-1β secretion. (E) Primary GEC were transfected with siRNA sequences against P2X 4 or P2X 7 for one day, and mRNA levels were detected by qPCR. (F) Primary GEC depleted of P2X 4 or P2X 7 were infected with P. gingivalis ( P.g. ) and treated with probenecid and 3 mM ATP as shown in (B). IL-1β secretion in the supernatants was analyzed by ELISA. The values showed averages and SD from duplicate samples, which were obtained from three separate experiments.

Article Snippet: The siRNA sequences were: 5′-GCUUUCAACGGGUCUGUCATT-3′ and 5′-UGACAGACCCGUUGAAAGCTA-3′ for P2X 4 (Ambion, LifeCell Technologies, S9957, Cat. #: 4392420); and 5′-ACAAUGUUGAGAAACGGACUCUGAT-3′ for P2X 7 (27 mer siRNA duplexes OriGene Technologies, Cat. #: SR303325).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Transfection

Journal: PLoS ONE

Article Title: P2X 4 Assembles with P2X 7 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: Model showing the role of P2X 4 and P2X 7 in ROS production and inflammasome activation in GEC stimulated with extracellular ATP.

Article Snippet: The siRNA sequences were: 5′-GCUUUCAACGGGUCUGUCATT-3′ and 5′-UGACAGACCCGUUGAAAGCTA-3′ for P2X 4 (Ambion, LifeCell Technologies, S9957, Cat. #: 4392420); and 5′-ACAAUGUUGAGAAACGGACUCUGAT-3′ for P2X 7 (27 mer siRNA duplexes OriGene Technologies, Cat. #: SR303325).

Techniques: Activation Assay