Journal: Translational Andrology and Urology

Article Title: Evaluation of transurethral GreenLight laser-selective vaporization for refractory overactive bladder in women

doi: 10.21037/tau-24-67

Figure Lengend Snippet: Expressions and comparisons of inflammatory factors and sensory receptors in bladder mucosa tissues before and after treatment. (A) The expressions of TRPV1, P2X3, TNF-α, and IL-6 in patients’ bladder mucosal tissues were stained by immunohistochemical staining before and at 12 weeks after treatment. Representative histological images are shown at 400× magnification. (B) Average optical density value of inflammatory factors and sensory receptors before and after treatment were presented in the bar chart. TNF, tumor necrosis factor; IL-6, interleukin 6; AOD, average optical density.

Article Snippet: Immunohistochemical staining was performed using the avidin-biotin-peroxidase method (ab64212, Abcam, Cambridge, MA, USA) with TRPV1 antibody (CSB-PA822774LA01HU, CUSABIO, Houston, TX, USA), P2X3 antibody (17843-1-AP, Proteintech, Rosemont, IL, USA), tumor necrosis factor alpha (TNF-α) antibody (17590-1-AP, Proteintech, Rosemont, IL, USA), and interleukin 6 (IL-6) antibody (21865-1-AP, Proteintech, Rosemont, IL, USA).

Techniques: Staining, Immunohistochemical staining

Journal: Acta neuropathologica communications

Article Title: Adenosine triphosphate drives head and neck cancer pain through P2X2/3 heterotrimers.

doi: 10.1186/2051-5960-2-62

Figure Lengend Snippet: Figure 1 ATP, P2X2, and P2X3 in human HNSCC microenvironment and pain. a. Representative HPLC chromatograms showing ATP peaks from tumor and matched normal tissue harvested from the same patient. b. Tumor tissues (Ipsi) had higher levels of ATP compared to matched normal sites (Contra) (n = 10, Student’s t-test). c. Pain scores of functional sharpness and intensity were significantly higher than spontaneous sharpness and intensity, respectively (n = 13, Student’s t-test). d. Mean scores of functional pain (Q2, 4, 6, 7, 8) were significantly higher than spontaneous pain (Q1, 3, 5) (Student’s t-test). e. ATP concentration in extracted cancer tissue correlated positively with mean pain scores (linear regression). f. Representative H&E and immunofluorescence staining (P2X2, P2X3, merged) of a human tongue SCC. Sections were taken from adjacent sections of the SCC. Scale bar: 100 μm.

Article Snippet: Control experiments were performed by incubation in secondary antibody alone and by applying P2X2 blocking peptides (Santa Cruz Biotechnology), and P2X3 blocking peptides (Alomone Labs).

Techniques: Functional Assay, Concentration Assay, Immunofluorescence, Staining

Journal: Acta neuropathologica communications

Article Title: Adenosine triphosphate drives head and neck cancer pain through P2X2/3 heterotrimers.

doi: 10.1186/2051-5960-2-62

Figure Lengend Snippet: Figure 3 HNSCC induces neuronal P2X2/3 plasticity that is reversed by anti-NGF. a. Sustained ATP current (top panel) is enhanced and prolonged following SCC co-culture; anti-NGF added into the co-culture reduced the sustained ATP current. Transient ATP current (lower panel) is not affected by either co-culture or anti-NGF. b. Sustained ATP current density is increased by co-culture (One-way ANOVA), and is reversed by anti-NGF. c. Dot plot of sustained ATP current in different diameter neurons. Co-culture increased current in medium-sized TG neurons; this increased current was reversed by anti-NGF. d. Representative immunofluorescence images of P2X2 and P2X3 expression in TG neurons. e. HNSCC co-culture increased the percentage of neurons expressing P2X2 and P2X3 subunits. Anti-NGF treatment significantly reduced percentage of neurons expressing P2X3 but not P2X2 subunits. The significant increase in the percentage of neurons that express both subunits following co-culture was reversed by anti-NGF application (One-way ANOVA). f. P2X2 immunofluorescence intensity was not changed after co-culture or anti-NGF treatment. P2X3 immunofluorescence intensity was significantly increased following co-culture, and was reduced by anti-NGF (one-way ANOVA). g. In mice with tongue HNSCC, mRNA expression for P2X2 was increased, while P2X3 expression was unchanged in TG neurons (Student’s t-test).

Article Snippet: Control experiments were performed by incubation in secondary antibody alone and by applying P2X2 blocking peptides (Santa Cruz Biotechnology), and P2X3 blocking peptides (Alomone Labs).

Techniques: Co-Culture Assay, Immunofluorescence, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Sensory Receptor, Inflammatory, and Apoptotic Protein Expression in the Bladder Urothelium of Patients with Different Subtypes of Interstitial Cystitis/Bladder Pain Syndrome

doi: 10.3390/ijms24010820

Figure Lengend Snippet: Western blots of bladder urothelial sensory receptors (TRPA1, TRPV1, TRPV4, P2X3, H-1, and sigma-1), and inflammatory (p38, tryptase) and pro-apoptotic proteins (caspase-3, BAX, BAD) in the bladder urothelium of IC/BPS patients and controls. More IC/BPS patients showed higher expression levels of TRPV1, TRPV4, sigma-1 receptors, P38, tryptase, caspase 3, and BAD expressions than controls. Molecular weight (kD) is indicated to the right of the blot. The GAPDH was blotted and used as a loading control.

Article Snippet: The slides were subsequently incubated with primary antibodies including TRPV1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX10296), TRPV4 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10296), TRPA1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX54765), P2X3 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10269), H-1 (dilution 1:25, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-374621), and sigma-1 (dilution 1:50, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-137075).

Techniques: Western Blot, Expressing, Molecular Weight, Control

Journal: International Journal of Molecular Sciences

Article Title: Sensory Receptor, Inflammatory, and Apoptotic Protein Expression in the Bladder Urothelium of Patients with Different Subtypes of Interstitial Cystitis/Bladder Pain Syndrome

doi: 10.3390/ijms24010820

Figure Lengend Snippet: The sensory, inflammatory, and pro-apoptotic protein expression levels in the urothelium of patients with interstitial cystitis/bladder pain syndrome and controls.

Article Snippet: The slides were subsequently incubated with primary antibodies including TRPV1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX10296), TRPV4 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10296), TRPA1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX54765), P2X3 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10269), H-1 (dilution 1:25, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-374621), and sigma-1 (dilution 1:50, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-137075).

Techniques: Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Sensory Receptor, Inflammatory, and Apoptotic Protein Expression in the Bladder Urothelium of Patients with Different Subtypes of Interstitial Cystitis/Bladder Pain Syndrome

doi: 10.3390/ijms24010820

Figure Lengend Snippet: The correlation between sensory receptor density and the clinical symptoms, inflammatory and pro-apoptotic protein expressions in the urothelium of patients with interstitial cystitis/bladder pain syndrome.

Article Snippet: The slides were subsequently incubated with primary antibodies including TRPV1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX10296), TRPV4 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10296), TRPA1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX54765), P2X3 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10269), H-1 (dilution 1:25, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-374621), and sigma-1 (dilution 1:50, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-137075).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Sensory Receptor, Inflammatory, and Apoptotic Protein Expression in the Bladder Urothelium of Patients with Different Subtypes of Interstitial Cystitis/Bladder Pain Syndrome

doi: 10.3390/ijms24010820

Figure Lengend Snippet: Sensory, inflammatory, and pro-apoptotic protein expression levels in the urothelium of patients with interstitial cystitis/bladder pain syndrome with different grades of glomerulations and controls.

Article Snippet: The slides were subsequently incubated with primary antibodies including TRPV1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX10296), TRPV4 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10296), TRPA1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX54765), P2X3 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10269), H-1 (dilution 1:25, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-374621), and sigma-1 (dilution 1:50, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-137075).

Techniques: Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Sensory Receptor, Inflammatory, and Apoptotic Protein Expression in the Bladder Urothelium of Patients with Different Subtypes of Interstitial Cystitis/Bladder Pain Syndrome

doi: 10.3390/ijms24010820

Figure Lengend Snippet: Sensory, inflammatory, and pro-apoptotic protein expression levels in the urothelium of patients with interstitial cystitis/bladder pain syndrome with maximal bladder capacity and controls.

Article Snippet: The slides were subsequently incubated with primary antibodies including TRPV1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX10296), TRPV4 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10296), TRPA1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX54765), P2X3 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10269), H-1 (dilution 1:25, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-374621), and sigma-1 (dilution 1:50, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-137075).

Techniques: Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Sensory Receptor, Inflammatory, and Apoptotic Protein Expression in the Bladder Urothelium of Patients with Different Subtypes of Interstitial Cystitis/Bladder Pain Syndrome

doi: 10.3390/ijms24010820

Figure Lengend Snippet: Sensory, inflammatory, and pro-apoptotic protein expression levels in the urothelium of patients with interstitial cystitis/bladder pain syndrome with different bladder pain severity and controls.

Article Snippet: The slides were subsequently incubated with primary antibodies including TRPV1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX10296), TRPV4 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10296), TRPA1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX54765), P2X3 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10269), H-1 (dilution 1:25, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-374621), and sigma-1 (dilution 1:50, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-137075).

Techniques: Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Sensory Receptor, Inflammatory, and Apoptotic Protein Expression in the Bladder Urothelium of Patients with Different Subtypes of Interstitial Cystitis/Bladder Pain Syndrome

doi: 10.3390/ijms24010820

Figure Lengend Snippet: Sensory, inflammatory, and pro-apoptotic protein expression levels in the urothelium of patients with interstitial cystitis/bladder pain syndrome with different bladder conditions during hydrodistention and controls.

Article Snippet: The slides were subsequently incubated with primary antibodies including TRPV1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX10296), TRPV4 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10296), TRPA1 (dilution 1:500, from GeneTex, CA, USA, catalog number: GTX54765), P2X3 (dilution 1:200, from GeneTex, CA, USA, catalog number: GTX10269), H-1 (dilution 1:25, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-374621), and sigma-1 (dilution 1:50, from Santa Cruz Biotechnology, CA, USA, catalog number: sc-137075).

Techniques: Expressing, Control

Journal: World Journal of Gastroenterology

Article Title: Study of the roles of caspase-3 and nuclear factor kappa B in myenteric neurons in a P2X7 receptor knockout mouse model of ulcerative colitis

doi: 10.3748/wjg.v29.i22.3440

Figure Lengend Snippet: Characteristics of primary and secondary antibodies

Article Snippet: P2X7 receptor , Rabbit , 1:200 , Merck Millipore.

Techniques:

Journal: World Journal of Gastroenterology

Article Title: Study of the roles of caspase-3 and nuclear factor kappa B in myenteric neurons in a P2X7 receptor knockout mouse model of ulcerative colitis

doi: 10.3748/wjg.v29.i22.3440

Figure Lengend Snippet: Colocalization of calretinin (red) with the P2X7 receptor (green) and DAPI (blue) in neurons in the myenteric plexus of the distal colon in mice. A1-A3: Wild-type (WT) sham 24 h; B1-B3: WT colitis 24 h; C1-C3: Knockout (KO) sham 24 h; D1-D3: KO colitis 24 h; E1-E3: WT sham 4 d; F1-F3: WT colitis 4 d; G1-G3: KO sham 4 d; H1-H3: KO colitis 4 d groups. The single arrows indicate calretinin-ir (A1-H1) and P2X7 receptor-ir (A2-H2) neurons, and the double arrows indicate the colocalization of calretinin with the P2X7 receptor and DAPI (A3-H3). Scale bar = 30 µm. WT: Wild-type; KO: Knockout.

Article Snippet: P2X7 receptor , Rabbit , 1:200 , Merck Millipore.

Techniques: Knock-Out

Journal: World Journal of Gastroenterology

Article Title: Study of the roles of caspase-3 and nuclear factor kappa B in myenteric neurons in a P2X7 receptor knockout mouse model of ulcerative colitis

doi: 10.3748/wjg.v29.i22.3440

Figure Lengend Snippet: Number of calretinin-ir and P2X7 receptor-ir neurons per myenteric ganglion in the distal colon in mice from the wild-type sham 24 h and 4 d, wild-type colitis 24 h and 4 d, knockout sham 24 h and 4 d, and knockout colitis 24 h and 4 d groups. A: Calretinin-ir; B: P2X7 receptor-ir. The data are presented as the arithmetic mean ± SE. a P < 0.05 vs sham groups; c P < 0.001 vs sham groups; NS: Not significant. WT: Wild-type; KO: Knockout.

Article Snippet: P2X7 receptor , Rabbit , 1:200 , Merck Millipore.

Techniques: Knock-Out

Journal: World Journal of Gastroenterology

Article Title: Study of the roles of caspase-3 and nuclear factor kappa B in myenteric neurons in a P2X7 receptor knockout mouse model of ulcerative colitis

doi: 10.3748/wjg.v29.i22.3440

Figure Lengend Snippet: Corrected total cell fluorescence of myenteric neurons. A: calretinin; B: P2X7 receptor; C: Cleaved caspase-3; D: Total caspase-3; E: Phospho-nuclear factor kappa B (NF-κB); F: Total NF-κB in the myenteric plexus of the distal colon in mice from the WT sham 24 h and 4 d, wild-type (WT) colitis 24 h and 4 d, knockout (KO) sham 24 h and 4 d, and KO colitis 24 h and 4 d groups. The data are presented as the arithmetic mean ± SE. a P < 0.05 vs sham groups; b P < 0.01 vs sham groups; c P < 0.001 vs sham groups; NS: Not significant. WT: Wild-type; KO: Knockout; NF-κB: Nuclear factor kappa B.

Article Snippet: P2X7 receptor , Rabbit , 1:200 , Merck Millipore.

Techniques: Fluorescence, Knock-Out