t1062  (TargetMol)

Journal: Research

Article Title: Na + /K + -ATPase Modulates Purinergic P2X3 Receptor Function to Drive Bone Cancer Pain

doi: 10.34133/research.0932

Figure Lengend Snippet: Na + /K + -ATPase α1 (NKAα1) interacts with P2X3 receptor (P2X3R) in dorsal root ganglion (DRG) neurons. (A) Quantitative polymerase chain reaction (qPCR) analysis showing the messenger RNA (mRNA) expression of P2rx1 , P2rx2 , P2rx3 , P2rx4 , P2rx5 , P2rx6 , and P2rx7 in the DRG ( n = 6). (B) Representative western blots and analyses of P2X3R in the DRG of sham and partial sciatic nerve ligation (PSNL) mice ( n = 4). (C) Representative western blots and analyses of P2X3R in the DRG of sham and tumor cell implantation (TCI) mice ( n = 4). (D) Annotated tandem mass spectrometry (MS/MS) spectra of 127-FPGGGILTGR-136 from P2X3R and 213-VDNSSLTGESEPQTR-227 from NKAα1 in anti-P2X3R immunoprecipitates from the DRG of 8-week naive mice. (E) Representative images and colocalization analyses of P2X3R (red) and NKAα1 (green) in DRG ( n = 8). The scale bars in (E) indicate 50 μm (left) and 10 μm (right, Zoom). (F and G) Co-immunoprecipitation analysis showing the interaction between NKAα1 and P2X3R in DRG. Data are presented as mean ± standard error of the mean (SEM). Statistical analysis by unpaired Student t test in (B) and (C). * P < 0.05; ** P < 0.01. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IP, immunoprecipitation; IgG, immunoglobulin G.

Article Snippet: To identify specific neuronal subtypes, β,γ-meATP (MCE, HY-134440A) was applied to mark P2X3R + cells, and capsaicin (TargetMol, USA, T1062) for TRPV1 + cells, and 50 mM KCl served as a positive control.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Ligation, Mass Spectrometry, Tandem Mass Spectroscopy, Immunoprecipitation

Journal: Research

Article Title: Na + /K + -ATPase Modulates Purinergic P2X3 Receptor Function to Drive Bone Cancer Pain

doi: 10.34133/research.0932

Figure Lengend Snippet: Impaired interaction between P2X3R and NKAα1 modulates P2X3R function in DRG neurons. (A to C) Representative images (A) and group data (B) showing Ca 2+ responses and traces of time-dependent intracellular Ca 2+ influx (C) induced by beta,gamma-methyleneadenosine 5′-triphosphate (β,γ-meATP) (100 μM) and capsaicin (1 μM) treatment. (D) Percentage of β,γ-meATP-responsive transient receptor potential vanilloid 1-positive (TRPV1 + ) neurons (defined as TRPV1 + neurons showing positive responses to capsaicin). Data represent 6 independent experiments per group, conducted on neurons derived from 2 independent cultures, with each experiment yielding 42 to 63 TRPV1 + responsive neurons. (E to G) Behavior tests showing the mechanical allodynia (E and F) and spontaneous pain-like behavior (G) after intraplantar injection of β,γ-meATP (10 nmol) in NKAα1 fl/fl and NKAα1 cKO mice ( n = 9 or 10). (H and I) Co-immunoprecipitation analysis showing that the interaction between NKAα1 and P2X3R in DRG was reduced in TCI mice ( n = 4). (J and K) Representative western blots and analyses of P2X3R and NKAα1 in the DRG of sham-NKAα1 fl/fl , TCI-NKAα1 fl/fl , sham-NKAα1 cKO , and TCI-NKAα1 cKO mice ( n = 3 or 4). The scale bars in (A) indicate 30 μm. Data are presented as mean ± SEM. Statistical analysis by unpaired Student t test in (B), (D), (F), (G), and (I) or 2-way ANOVA with Bonferroni’s post hoc test in (E) and (K). NS, not significant. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: To identify specific neuronal subtypes, β,γ-meATP (MCE, HY-134440A) was applied to mark P2X3R + cells, and capsaicin (TargetMol, USA, T1062) for TRPV1 + cells, and 50 mM KCl served as a positive control.

Techniques: Derivative Assay, Injection, Immunoprecipitation, Western Blot

Journal: Research

Article Title: Na + /K + -ATPase Modulates Purinergic P2X3 Receptor Function to Drive Bone Cancer Pain

doi: 10.34133/research.0932

Figure Lengend Snippet: Schematic diagram illustrating the proposed mechanism in the TCI model: injury-induced activation of P2X3R channels leads to Ca 2+ influx, which down-regulates protein phosphatase 2A (PP2A) expression and diminishes membrane localization of NKAα1. The decreased membrane NKAα1 disrupts its interaction with P2X3R, further promoting Ca 2+ influx and additional suppression of PP2A. Concurrently, reduced PP2A expression activates the extracellular signal-regulated kinase 1/2 (ERK1/2)–Runt-related transcription factor 1 (Runx1) signaling pathway, upregulating P2X3R expression. Together, these changes enhance the excitability of neurons and neuronal injury, which in turn promotes CCL5 release and glial cell activation, ultimately inducing bone cancer pain. SDH, spinal dorsal horn.

Article Snippet: To identify specific neuronal subtypes, β,γ-meATP (MCE, HY-134440A) was applied to mark P2X3R + cells, and capsaicin (TargetMol, USA, T1062) for TRPV1 + cells, and 50 mM KCl served as a positive control.

Techniques: Activation Assay, Expressing, Membrane

Journal: Frontiers in Pharmacology

Article Title: Long-term nitric oxide exposure induces cough hypersensitivity via non-inflammatory activation of the HIF1α–TRPV1 pathway

doi: 10.3389/fphar.2026.1679727

Figure Lengend Snippet: NO stimulation does not induce coughing, and long-term NO exposure increases cough sensitivity in guinea pigs. (A) Cough detection setup for guinea pigs after capsaicin and SNP nebulization. (B) Cough frequency in guinea pigs after capsaicin and SNP nebulization ( n = 5; mean ± SEM; ** P = 0.0016; two-tailed unpaired Student’s t-test). (C) NO exposure chamber setup for guinea pigs. (D) Cough frequency in guinea pigs recorded after 4 weeks of NO exposure, following capsaicin stimulation ( n = 6; mean ± SEM; ** P = 0.0012; two-tailed unpaired Student’s t-test). (E) Latency to the first cough in guinea pigs after 4 weeks of NO exposure, recorded following capsaicin stimulation ( n = 6; mean ± SEM; * P = 0.0493; two-tailed unpaired Student’s t-test).

Article Snippet: Cough challenge tests were performed using a BUXCO system (Buxco, Data Sciences International, New York, United States) following aerosol stimulation with capsaicin (CAP, TargetMol, Boston, MA, United States) or sodium nitroprusside (SNP, Beyotime, Shanghai, China).

Techniques: Two Tailed Test