Fig 1: Postsynaptic density protein-95 (PSD-95) forms a complex with NR2B-subtype receptors in dorsal horn neurons and binding to NR2B subunits is perturbed by Tat-NR2B9c. (a) Western immunoblot showing expression of PSD-95 in hippocampus (two left lanes), dorsal root ganglion (DRG) (two centre lanes), and spinal dorsal horn (two right lanes). ßIII-tubulin served as a loading control. (b) Western immunoblots of normal lumbar spinal dorsal horn lysates (left two lanes) and coimmunoprecipitates from lumbar spinal dorsal horn lysate obtained using an antibody against NR2B (right two lanes), probed with antibodies against NR2B, neuronal nitric oxide synthase (nNOS), PSD-95, CREB, and P2X3. (c) Western immunoblots showing immunoprecipitates (IP, right lanes) from the lumbar dorsal horn of rats pretreated intrathecally with Tat-NR2BAA (125 ng) or Tat-NR2B9c (125 ng), 20 minutes before being sacrificed, obtained using an antibody against NR2B subunits (representative of four independent experiments). Note the marked reduction in PSD-95 coimmunoprecipitated with NR2B subunits from the dorsal horn of rats pretreated with Tat-NR2B9c. Left lane shows normal dorsal horn lysates with no immunoprecipitation, acting as positive controls.
Fig 2: P2X3-immunoreactive staining in endometriosis endometrium and endometriotic lesions as compared with control endometrium.(A), Control endometrium from a woman without endometriosis; (B), Endometriosis endometrium from a woman with ovarian endometriosis; (C), Ovarian endometriotic lesions from a woman with ovarian endometriosis. The immunohistochemistry (IHC) score of P2X3 expression in endometriosis endometrium and endometriotic lesions were both significantly higher as compared with control endometrium (P<0.05; Original magnification 400×; Bar = 50 µm).
Fig 3: P2X3 protein and mRNA expression in the colonic myenteric plexus. (a) Distribution of the P2X3 receptor in the colonic myenteric plexus (scale bar =20 µm). (b) P2X3 mRNA expression in colon. *P < 0.05, **P < 0.01.
Fig 4: The changed levels of p-ERK, p-CREB, and P2X3 expression in ESCs after treated with ATP and ERK inhibitor.(A, B, C and D), The ESCs were treated with ATP alone; (E, F, G and H), The ESCs were pretreated with ERK inhibitor for 45min, and then treated with ATP. ESCs: Endometriotic stromal cells. A. The levels of P2X3 mRNA expression in ESCS increased at 15min, 30 min and 1h after treated with ATP. B. Western blot showed a specific band (47 kDa) for P2X3, and the levels of P2X3 expression in ESCs increased at 15min, continued to increase at 30min, reached the peak value at 1h, and then gradually decreased at 2h. C. Western blot showed two bands (42/44 kDa) for p-ERK, and the levels of p-ERK expression in ESCs reached the peak value at 15min, and then gradually decreased at 30min. D. Western blot showed a specific band (45 kDa) for p-CREB, and the levels of p-CREB expression in ESCs increased at 15min, reached the peak value at 30min, and then gradually decreased at 1h. E. The elevated levels of P2X3 mRNA induced by ATP were totally blocked by ERK1/2 inhibitor. F. The levels of P2X3 protein expression in ESCs did not increase at any time points. G. The levels of p-ERK expression in ESCs significantly decreased at any time points. H. The levels of p-CREB expression in ESCs at any time points except at 1h did not increase. (* P<0.05. **P<0.01. ***P<0.00001.)
Fig 5: Correlation of VAS score and P2X3 expression IHC score in endometriosis endometrium and endometriotic lesions.A. endometriosis endometrium; B. endometriotic lesions. (VAS), Visual analog scale; (IHC), Immunohistochemical staining. The IHC scores of P2X3 expression in endometriosis endometrium and endometriotic lesions were both correlated with VAS score in women with endometriosis (P<0.05).
Supplier Page from Abcam for Anti-P2X3 antibody