Journal: Hypertension research : official journal of the Japanese Society of Hypertension

Article Title: P2X(7) receptor antagonism attenuates the hypertension and renal injury in Dahl salt-sensitive rats.

doi: 10.1038/hr.2011.153

Figure Lengend Snippet: Figure 1 P2X7 expression in the kidneys of DS and LEW rats. (a) P2X7 mRNA levels in the kidneys of the DS and LEW rats. Real-time RT-PCR analyses were performed using GAPDH mRNA levels as an internal control. The figure shows a summary of the data analyzed by analysis of variance (ANOVA) (N¼6 for each group). Multiple regression analysis indicated that P2X7 mRNA levels in the kidneys were significantly influenced by the difference in the strains (DS or LEW) and by the amount of salt in the diet (high- or normal-salt diet). P2X7 mRNA levels in the kidneys of the DS rats were significantly higher than those in the kidneys of the LEW rats (DS vs. LEW, Po0.0001, by ANOVA). (b) P2X7 protein expression in the kidneys of the DS and LEW rats. Western blot analysis indicated that P2X7 protein levels in the kidneys of the DS rats were significantly higher than those in the kidneys of the LEW rats (N¼6 for each group) (DS vs. LEW, Po0.0001, by ANOVA).

Article Snippet: After blocking with 5% (w/v) skim milk powder, membranes were incubated with the primary rat P2X7 antibody (APR-008; Alomone Labs, Jerusalem, Israel) at 1:1000 dilution, and the membrane-bound antibody was visualized with horseradish peroxidase- conjugated secondary antibody (NA934V; Amersham Biopharmacia, Buckin- ghamshire, UK), diluted 1:10 000, and an ECL advance western blotting detection kit (GE Healthcare). b-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used as a loading control.

Techniques: Expressing, Quantitative RT-PCR, Control, Western Blot

Journal: Hypertension research : official journal of the Japanese Society of Hypertension

Article Title: P2X(7) receptor antagonism attenuates the hypertension and renal injury in Dahl salt-sensitive rats.

doi: 10.1038/hr.2011.153

Figure Lengend Snippet: Figure 2 Blockade of P2X7 in the DS rats with BBG. (a) Effects on blood pressure. In all 4 weeks of the 8% NaCl diet significantly increased blood pressure in the DS rats (N¼7 for both groups). Multivariate analysis of variance (MANOVA) indicated that BBG treatment significantly attenuated this rise in blood pressure (Po0.0001). *Po0.01, significantly different from the values obtained in the 8% NaCl diet group (Student’s t-test). (b) Effects on renal weight. Kidneys were harvested at 4 weeks with or without BBG treatment (N¼7 for both groups), and renal weight was represented as a relative weight (renal organ weight per 10 g of body weight). Renal weight in the DS rats without BBG treatment was significantly higher than in those with BBG treatment (Po0.005, Student’s t-test). (c) Effects on urinary protein excretion. A total of 4 weeks of the 8% NaCl diet significantly increased urinary protein excretion in the DS rats (N¼7 for both groups). MANOVA indicated that BBG treatment significantly attenuated this increase in urinary protein excretion (P¼0.0006). *Po0.05, **Po0.01, significantly different from the values obtained in the 8% NaCl diet group (Student’s t-test). (d) Effects on creatinine clearance. Creatinine clearance (CCr) at 4 weeks was measured, which was calculated using the formula: CCr¼(UCrV)/PCr, where UCr is the concentration of urinary creatinine (mg dl1), PCr is the concentration of plasma creatinine (mgdl1), and V is the urine flow rate (ml min1). Creatinine clearance significantly improved by 4 weeks of BBG treatment (Po0.001, Student’s t-test).

Article Snippet: After blocking with 5% (w/v) skim milk powder, membranes were incubated with the primary rat P2X7 antibody (APR-008; Alomone Labs, Jerusalem, Israel) at 1:1000 dilution, and the membrane-bound antibody was visualized with horseradish peroxidase- conjugated secondary antibody (NA934V; Amersham Biopharmacia, Buckin- ghamshire, UK), diluted 1:10 000, and an ECL advance western blotting detection kit (GE Healthcare). b-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used as a loading control.

Techniques: Concentration Assay, Clinical Proteomics

Journal: Hypertension research : official journal of the Japanese Society of Hypertension

Article Title: P2X(7) receptor antagonism attenuates the hypertension and renal injury in Dahl salt-sensitive rats.

doi: 10.1038/hr.2011.153

Figure Lengend Snippet: Figure 6 P2X7 in peritoneal macrophages from the DS and LEW rats without salt loading. (a) P2X7 mRNA levels in the peritoneal macrophages from the DS and LEW rats. Real-time RT-PCR analyses were performed using GAPDH mRNA levels as an internal control. The figure shows a summary of the data analyzed by ANOVA (N¼8 for each group). P2X7 mRNA levels in the macrophages from the DS rats were significantly higher than those in the macrophages from the LEW rats (DS vs. LEW, Po0.0001). (b) IL-1b release from the LPS-primed macrophages. Peritoneal macrophages were isolated from the DS rats (N¼13) and LEW rats (N¼13). LPS-primed macrophages were subjected to BzATP stimulation (100 mM, for 6 h at 371C) for the assessment of IL-1b release. Significantly higher release of IL-1b was observed from the macrophages of the DS rats compared with that from the macrophages of the LEW rats (P¼0.0002).

Article Snippet: After blocking with 5% (w/v) skim milk powder, membranes were incubated with the primary rat P2X7 antibody (APR-008; Alomone Labs, Jerusalem, Israel) at 1:1000 dilution, and the membrane-bound antibody was visualized with horseradish peroxidase- conjugated secondary antibody (NA934V; Amersham Biopharmacia, Buckin- ghamshire, UK), diluted 1:10 000, and an ECL advance western blotting detection kit (GE Healthcare). b-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used as a loading control.

Techniques: Quantitative RT-PCR, Control, Isolation

Journal: Hypertension research : official journal of the Japanese Society of Hypertension

Article Title: P2X(7) receptor antagonism attenuates the hypertension and renal injury in Dahl salt-sensitive rats.

doi: 10.1038/hr.2011.153

Figure Lengend Snippet: Figure 7 Co-localization of P2X7 and CD68 (macrophage marker) in renal consecutive sections. On immunohistochemical examination, P2X7 and macrophages were identified by positive staining with anti-P2X7 and anti- CD68 antibodies, respectively. It is shown that macrophages are co- expressed with P2X7 (arrowheads). Lower panels are magnified photographs of upper ones (rectangular area). Scale bars¼50mm.

Article Snippet: After blocking with 5% (w/v) skim milk powder, membranes were incubated with the primary rat P2X7 antibody (APR-008; Alomone Labs, Jerusalem, Israel) at 1:1000 dilution, and the membrane-bound antibody was visualized with horseradish peroxidase- conjugated secondary antibody (NA934V; Amersham Biopharmacia, Buckin- ghamshire, UK), diluted 1:10 000, and an ECL advance western blotting detection kit (GE Healthcare). b-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used as a loading control.

Techniques: Marker, Immunohistochemical staining, Staining

Journal: iScience

Article Title: Hepatitis B virus-mediated sodium influx contributes to hepatic inflammation via synergism with intrahepatic danger signals

doi: 10.1016/j.isci.2023.108723

Figure Lengend Snippet:

Article Snippet: anti-P2X7 , Cell signaling Techonology , 13809.

Techniques: Activity Assay, ATP Assay, Enzyme-linked Immunosorbent Assay, Sequencing, Plasmid Preparation, RNA Sequencing Assay, Biomarker Assay, Software

Journal: Journal of advanced research

Article Title: The purinergic receptor P2rx7 mediated ATP sensing is required to prevent bone aging by directing mitochondrial fitness of MSCs.

doi: 10.1016/j.jare.2025.06.026

Figure Lengend Snippet: Fig. 2. P2rx7 promoted the bone formation of MSCs. a The CFU-F in MSCs derived from the control and the P2rx7-/- MSCs. b Mineralized nodules formed by MSCs of the control and the P2rx7-/- mice were analyzed by alizarin red staining. c, d The expressions of osteogenesis marker genes in the control and the P2rx7-/- MSCs, as assessed by western blotting (c) and qPCR (d). e, f Transplants consisting of MSCs from control and P2rx7-/- mice mixed with HA/TCP were analyzed by H&E staining (e), and immunofluorescent staining of CD31 and ACAN (f), n = 5 per group. g Mineralized nodules formed by MSCs under osteogenic induction when exposed to 0, 40 nM or 100 nM BzATP were showed by alizarin red staining. h The expressions of osteogenic markers Runx2, ALP, and OCN in MSCs exposed to 0, 40 nM or 100 nM BzATP, as assessed by western blotting. i, j Transplants consisting of MSCs treated with 0, 40, 100 nM BzATP from control mice mixed with HA/TCP were analyzed by H&E staining (i), and immunofluorescent staining of CD31 and ACAN (j), n = 5 per group. Scale bar: e, f, i, j: 50 lm. e, i yellow, new bone; black, blood vessel; blue, new cartilage. Data was presented as mean ± SD. The data (a-d, g) are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs con by two-tailed Student’s t test (a, b, e, f) and one-way ANOVA (d, g, i, j). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The antibodies used in this study were as follows: Anti-mouse CD29 (#2473836), anti-mouse CD31 (#2373735), anti-mouse CD44 (#2255545) antibodies from eBioscience (San Diego, CA, USA); PE anti-mouse CD11b (#101207), APC anti-mouse CD90.2 (#105311) antibodies from BioLegend (San Diego, CA, USA); Anti-Col1a1 (ab255809; Abcam, Cambridge, MA, USA); Drp1 (#8570), Erk1/2 (#4370), p-Erk1/2 (#4695) from CST (Danvers, MA, USA); alkaline phosphatase (ALP, PA5-106391; Invitrogen, Carlsbad, CA, USA); Bmp-2 (sc-137087), CD90 (sc-53116), Cytoc (sc-13156), Ocn (sc-390877), P2rx7 (sc-514962) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Fis1 (10956–1-AP), p16 (10883–1-AP), p21 (10355–1-AP), P2rx7 (28207–1-AP), Mfn1 (13798–1-AP), Mfn2 (12186–1-AP), Opa1 (27733–1-AP), Runx2 (82636–2-RR) from ProteinTech (Cook, Illinois, USA); p-Drp1 (Ser616) (AF8470) from Affinity (Liyang, China); Hsp60 (AF0186) and Mouse IgG (A7028) from Beyotime (Shanghai, China); bActin (GB15001-100) from Servicebio (Wuhan, China).

Techniques: Derivative Assay, Control, Staining, Marker, Western Blot, Two Tailed Test

Journal: Journal of advanced research

Article Title: The purinergic receptor P2rx7 mediated ATP sensing is required to prevent bone aging by directing mitochondrial fitness of MSCs.

doi: 10.1016/j.jare.2025.06.026

Figure Lengend Snippet: Fig. 3. P2rx7 controls mitochondrial dynamics in MSCs. a OCR, calculated basal respiration and ATP-linked respiration of the control and the P2rx7-/- MSCs. b ATP contents of the control and the P2rx7-/- MSCs. c ECAR, and calculated glycolysis, glycolytic capability and glycolytic reverse of the control and the P2rx7-/- MSCs. d TEM images of mitochondria (arrows) in the control and the P2rx7-/- MSCs (n = 20 per group). e DWM of the control and the P2rx7-/- MSCs was measured by JC-1 staining (polymer: red; monomer: green). f The expressions of genes Nd1 and mt-Cytb in the control and the P2rx7-/- MSCs that related to the mitochondrial mass in cells assessed by qPCR. g DWM of MSCs treated with different concentrations of BzATP was analyzed by JC-1 staining. Scale bar: d: 500 nm; e, g: 100 lm. Data was presented as mean ± SD. The data (a, b, e-g) are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs con by two-tailed Student’s t test (a-f) and one-way ANOVA (g). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The antibodies used in this study were as follows: Anti-mouse CD29 (#2473836), anti-mouse CD31 (#2373735), anti-mouse CD44 (#2255545) antibodies from eBioscience (San Diego, CA, USA); PE anti-mouse CD11b (#101207), APC anti-mouse CD90.2 (#105311) antibodies from BioLegend (San Diego, CA, USA); Anti-Col1a1 (ab255809; Abcam, Cambridge, MA, USA); Drp1 (#8570), Erk1/2 (#4370), p-Erk1/2 (#4695) from CST (Danvers, MA, USA); alkaline phosphatase (ALP, PA5-106391; Invitrogen, Carlsbad, CA, USA); Bmp-2 (sc-137087), CD90 (sc-53116), Cytoc (sc-13156), Ocn (sc-390877), P2rx7 (sc-514962) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Fis1 (10956–1-AP), p16 (10883–1-AP), p21 (10355–1-AP), P2rx7 (28207–1-AP), Mfn1 (13798–1-AP), Mfn2 (12186–1-AP), Opa1 (27733–1-AP), Runx2 (82636–2-RR) from ProteinTech (Cook, Illinois, USA); p-Drp1 (Ser616) (AF8470) from Affinity (Liyang, China); Hsp60 (AF0186) and Mouse IgG (A7028) from Beyotime (Shanghai, China); bActin (GB15001-100) from Servicebio (Wuhan, China).

Techniques: Control, Staining, Polymer, Two Tailed Test

Journal: Journal of advanced research

Article Title: The purinergic receptor P2rx7 mediated ATP sensing is required to prevent bone aging by directing mitochondrial fitness of MSCs.

doi: 10.1016/j.jare.2025.06.026

Figure Lengend Snippet: Fig. 4. P2rx7 regulated mitochondrial fusion through Mfn1. a The mitochondria (top) labeled by Mitotracker Red and mitochondrial morphology (bottom) based on Mitochondrial Analyzer after grey processing, analyzed by Airyscan confocal images in the control and P2rx7-/- MSCs. b The expression of genes associated with mitochondrial dynamics in the control and the P2rx7-/- MSCs as assessed by western blotting. c Representative micrographs visualizing P2rx7 (arrows) on the mitochondria. d Reprehensive Airyscan confocal images of mitochondrial morphology (Mitotracker Red, top) and mitochondrial morphology (bottom) based on Mitochondrial Analyzer after grey processing in the MSCs treated with 0, 40 nM and 100 nM BzATP. e The expression of genes associated with mitochondrial dynamics in the MSCs treated with 0, 40 nM and 100 nM BzATP as assessed by western blotting. f Western blotting analysis of mitochondrial (left) and cytosolic (right) fractions of the MSCs treated with 40 nM and 100 nM BzATP. g IHC staining of Mfn1 and Opa1 proteins in the control and the P2rx7-/- MSCs that were subcutaneously implanted into immunocompromised mice with HA/TCP (n = 5 per group). Scale bar: a, d:10 lm (top), 5 lm(bottom); g: 100 lm; h: 500 lm. Data was presented as mean ± SD. The data (a, b, d-f) are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs con by two-tailed Student’s t test. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The antibodies used in this study were as follows: Anti-mouse CD29 (#2473836), anti-mouse CD31 (#2373735), anti-mouse CD44 (#2255545) antibodies from eBioscience (San Diego, CA, USA); PE anti-mouse CD11b (#101207), APC anti-mouse CD90.2 (#105311) antibodies from BioLegend (San Diego, CA, USA); Anti-Col1a1 (ab255809; Abcam, Cambridge, MA, USA); Drp1 (#8570), Erk1/2 (#4370), p-Erk1/2 (#4695) from CST (Danvers, MA, USA); alkaline phosphatase (ALP, PA5-106391; Invitrogen, Carlsbad, CA, USA); Bmp-2 (sc-137087), CD90 (sc-53116), Cytoc (sc-13156), Ocn (sc-390877), P2rx7 (sc-514962) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Fis1 (10956–1-AP), p16 (10883–1-AP), p21 (10355–1-AP), P2rx7 (28207–1-AP), Mfn1 (13798–1-AP), Mfn2 (12186–1-AP), Opa1 (27733–1-AP), Runx2 (82636–2-RR) from ProteinTech (Cook, Illinois, USA); p-Drp1 (Ser616) (AF8470) from Affinity (Liyang, China); Hsp60 (AF0186) and Mouse IgG (A7028) from Beyotime (Shanghai, China); bActin (GB15001-100) from Servicebio (Wuhan, China).

Techniques: Labeling, Control, Expressing, Western Blot, Immunohistochemistry, Two Tailed Test

Journal: Journal of advanced research

Article Title: The purinergic receptor P2rx7 mediated ATP sensing is required to prevent bone aging by directing mitochondrial fitness of MSCs.

doi: 10.1016/j.jare.2025.06.026

Figure Lengend Snippet: Fig. 5. P2rx7 enhanced the sensitivity of ERK pathway to promote mitochondrial fusion. a Top 10 significantly enrichment of downregulated DEGs between control and P2rx7-/- MSCs analyzed by KEGG. b Expressions of Erk1/2 and p-Erk1/2 protein in the distal femurs were analyzed by IHC staining (n = 5 per group). c The phosphorylation level and the total expression level of Erk1/2 in the control and the P2rx7-/- MSCs before and after the activated by bFGF, as assessed by western blotting. d The phosphorylation level and the total expression level of Erk1/2 in MSCs treated with different concentrations of BzATP before and after the activated by 10 lM bFGF for 1 h, as assessed by western blotting. e Expressions of genes associated with mitochondrial dynamics in the Widetype (WT) and the P2rx7-/- MSCs treated with bFGF as assessed by western blotting. f Expressions of genes associated with mitochondrial dynamics in MSCs treated with siNC, siMfn1 and BzATP as assessed by western blotting. g DWM of the Widetype (WT) and the P2rx7-/-MSCs treated with BzATP and bFGF was analyzed by JC-1 staining. h ALP activity (top) and mineralized nodules (bottom) under osteogenic induction of human MSCs when exposed to 100 nM BzATP in the presence and absence of U0126. i The expression of osteogenesis marker genes in MSCs exposed with BzATP, in the presence and absence of 10 lM U0126. Scale bar: b: 100 lm; d: 50 lm; h: 500 lm. Data was presented as mean ± SD. The data (c-i) are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs con by two-tailed Student’s t test (b) and one-way ANOVA (g, h).

Article Snippet: The antibodies used in this study were as follows: Anti-mouse CD29 (#2473836), anti-mouse CD31 (#2373735), anti-mouse CD44 (#2255545) antibodies from eBioscience (San Diego, CA, USA); PE anti-mouse CD11b (#101207), APC anti-mouse CD90.2 (#105311) antibodies from BioLegend (San Diego, CA, USA); Anti-Col1a1 (ab255809; Abcam, Cambridge, MA, USA); Drp1 (#8570), Erk1/2 (#4370), p-Erk1/2 (#4695) from CST (Danvers, MA, USA); alkaline phosphatase (ALP, PA5-106391; Invitrogen, Carlsbad, CA, USA); Bmp-2 (sc-137087), CD90 (sc-53116), Cytoc (sc-13156), Ocn (sc-390877), P2rx7 (sc-514962) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Fis1 (10956–1-AP), p16 (10883–1-AP), p21 (10355–1-AP), P2rx7 (28207–1-AP), Mfn1 (13798–1-AP), Mfn2 (12186–1-AP), Opa1 (27733–1-AP), Runx2 (82636–2-RR) from ProteinTech (Cook, Illinois, USA); p-Drp1 (Ser616) (AF8470) from Affinity (Liyang, China); Hsp60 (AF0186) and Mouse IgG (A7028) from Beyotime (Shanghai, China); bActin (GB15001-100) from Servicebio (Wuhan, China).

Techniques: Control, Immunohistochemistry, Phospho-proteomics, Expressing, Western Blot, Staining, Activity Assay, Marker, Two Tailed Test

Journal: Journal of advanced research

Article Title: The purinergic receptor P2rx7 mediated ATP sensing is required to prevent bone aging by directing mitochondrial fitness of MSCs.

doi: 10.1016/j.jare.2025.06.026

Figure Lengend Snippet: Fig. 6. Bone metabolism and regeneration could be promoted by DCA treatment. a, b Increased mineralized nodules (a) and ALP activity (b) under osteogenic induction of the P2rx7-/- MSCs treated with PBS (Control) and 5 lM DCA. c, d Expressions of osteogenesis marker genes in the P2rx7-/- MSCs treated with Control and 5 lM DCA, as assessed by western blotting (c) and qPCR analysis (d). e, f Cross-sectional lCT images of the trabecular bone (e) and H&E staining of distal femurs (f) of 4-month-old P2rx7-/- mice treated with and without DCA, n = 5 per group. g, h Mouse mandibles of the Control and the BzATP group after 4 weeks of healing were presented as lCT images (g) and statistical analysis (g right, BV/TV in lCT images; h right, percent of area of bone tissue according to Masson staining), H&E staining (h, top) and Masson staining (h, bottom), n = 5 per group. Black arrow: new bone. i Expressions of Mfn1 and Opa1 proteins in the area of defect after 4-week healing were analyzed by IHC staining (n = 5 per group). Scale bar: f, h: 500 lm; i: 100 lm. Data was presented as mean ± SD. The data (a-d) are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs con by two- tailed Student’s t test.

Article Snippet: The antibodies used in this study were as follows: Anti-mouse CD29 (#2473836), anti-mouse CD31 (#2373735), anti-mouse CD44 (#2255545) antibodies from eBioscience (San Diego, CA, USA); PE anti-mouse CD11b (#101207), APC anti-mouse CD90.2 (#105311) antibodies from BioLegend (San Diego, CA, USA); Anti-Col1a1 (ab255809; Abcam, Cambridge, MA, USA); Drp1 (#8570), Erk1/2 (#4370), p-Erk1/2 (#4695) from CST (Danvers, MA, USA); alkaline phosphatase (ALP, PA5-106391; Invitrogen, Carlsbad, CA, USA); Bmp-2 (sc-137087), CD90 (sc-53116), Cytoc (sc-13156), Ocn (sc-390877), P2rx7 (sc-514962) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Fis1 (10956–1-AP), p16 (10883–1-AP), p21 (10355–1-AP), P2rx7 (28207–1-AP), Mfn1 (13798–1-AP), Mfn2 (12186–1-AP), Opa1 (27733–1-AP), Runx2 (82636–2-RR) from ProteinTech (Cook, Illinois, USA); p-Drp1 (Ser616) (AF8470) from Affinity (Liyang, China); Hsp60 (AF0186) and Mouse IgG (A7028) from Beyotime (Shanghai, China); bActin (GB15001-100) from Servicebio (Wuhan, China).

Techniques: Activity Assay, Control, Marker, Western Blot, Staining, Immunohistochemistry, Two Tailed Test

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