Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Molecular mechanisms of voiding dysfunction in a novel mouse model of acute urinary retention

doi: 10.1096/fj.202002415R

Figure Lengend Snippet: Altered receptor expression in mediating diminished BSM contractility upon pressure treatment. A, Western blot images of M2, M3, and P2X1 receptor protein expression in mouse bladders from control (n = 4) and pressure treated mice (50 cm: n = 4 for 0 hours, 6 hours, and 24 hours, respectively. 80 cm, n = 4 for 0 hours, 6 hours, and 24 hours, respectively). B-D, are quantitated data by densitometry and normalized to actin. Data are shown as box and whiskers, center line is the median of the data set, box represents 75% of the data, and bars indicate whiskers from minimum to maximum. Data were analyzed by Student’s t test between control group and 50 or 80 cm H2O pressure treated groups. *P < .05, **P < .001

Article Snippet: Western blot analysis Rabbit anti-P2X1 antibody (1:1000, #APR-001) and rabbit anti-chrm2 antibody (1:1000, #AMR-002) were purchased from Alomone Lab. Rabbit anti-chrm3 antibody (1:1000, #PA5–77485) was purchased from Thermo Fisher Scientific.

Techniques: Expressing, Western Blot

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: (A) A schematic diagram of the maximum cross section of a rat nodose ganglion (A, corresponding to panel B). (B) Representative graph of the distribution of P2X receptor-positive cells throughout the whole nodose ganglion section under a 20× light microscopic field. (C–F) Immunohistochemical staining using polyclonal antibodies against P2X1 (C), P2X2 (D), P2X3 (E), and P2X4 (F); 100× magnification. Scale bars in B = 500 µm and F = 100 µm.

Article Snippet: Samples were blocked with donkey serum for 1 h. Next, samples were incubated with rabbit anti-P2X1, mouse anti-P2X2, goat anti-P2X3, and chicken anti-P2X4 polyclonal antibodies (1∶200, Santa Cruz Biotechnology, Santa Cruz, CA) for 48 h. FITC- and TRITC- conjugated secondary antibodies were then incubated for 1 h.

Techniques: Immunohistochemical staining, Staining

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: (A), (B), (C), and (D) show P2X–IR sections that were counterstained with neutral red to precisely calculate the number of P2X1, P2X2, P2X3, and P2X4-positive neurons, respectively (using black positive cells compared with red negative cells). Scale bar in (D) = 100 µm. (E), (F), (G), and (H) represent the frequency distribution of the area-size of P2X1, P2X2, P2X3, and P2X4-IR, respectively.

Article Snippet: Samples were blocked with donkey serum for 1 h. Next, samples were incubated with rabbit anti-P2X1, mouse anti-P2X2, goat anti-P2X3, and chicken anti-P2X4 polyclonal antibodies (1∶200, Santa Cruz Biotechnology, Santa Cruz, CA) for 48 h. FITC- and TRITC- conjugated secondary antibodies were then incubated for 1 h.

Techniques:

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: The upper panel shows immunohistochemical staining for P2X1–4 subunits in the maximum cross section of the NG tissues. The lower panel shows that P2X3-IR was the most prevalent neuronal subtype, following by P2X2-IR; P2X4-IR had the lowest prevalence. * p <0.05.

Article Snippet: Samples were blocked with donkey serum for 1 h. Next, samples were incubated with rabbit anti-P2X1, mouse anti-P2X2, goat anti-P2X3, and chicken anti-P2X4 polyclonal antibodies (1∶200, Santa Cruz Biotechnology, Santa Cruz, CA) for 48 h. FITC- and TRITC- conjugated secondary antibodies were then incubated for 1 h.

Techniques: Immunohistochemical staining, Staining

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: NG neurons were stained using antibodies against P2X1, P2X2, P2X3, and P2X4 receptor subunits (A, D, G, and J, respectively). The nuclei of cultured NG neurons were stained with antibodies against NeuN (B, E, H, K). Merged images (C, F, I, L) representing co-staining of P2X receptor subunits and NeuN are shown. The scale bar shown in L is representative of all images, and represents 50 µm.

Article Snippet: Samples were blocked with donkey serum for 1 h. Next, samples were incubated with rabbit anti-P2X1, mouse anti-P2X2, goat anti-P2X3, and chicken anti-P2X4 polyclonal antibodies (1∶200, Santa Cruz Biotechnology, Santa Cruz, CA) for 48 h. FITC- and TRITC- conjugated secondary antibodies were then incubated for 1 h.

Techniques: Staining, Cell Culture

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: (A–C) Immunoreactivity of P2X1 and P2X2 (A–C), P2X1 and P2X3 (D–F), and P2X2 and P2X3 (G–I) subunits in the NG. Scale bars = 50 µm.

Article Snippet: Samples were blocked with donkey serum for 1 h. Next, samples were incubated with rabbit anti-P2X1, mouse anti-P2X2, goat anti-P2X3, and chicken anti-P2X4 polyclonal antibodies (1∶200, Santa Cruz Biotechnology, Santa Cruz, CA) for 48 h. FITC- and TRITC- conjugated secondary antibodies were then incubated for 1 h.

Techniques:

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: (A) Schematic view of the setup for the whole cell patch clamp and a representative image of a recorded cell under the phase contrast microscope and immunohistochemistry. (B) Immunohistochemistry revealed positive or negative staining for P2X1–4 subunits, which correlated with the type of I ATP and cell size. The samples in each row were from four different neurons that responded to ATP with different types of ATP-activated current. P2X3 staining was positive in all four types of I ATP neurons. P2X1 was positive in F, I, and S I ATP s, but negative in VS. P2X2 staining was only absent in neurons with type I I ATP , and P2X4 was positive in neurons with type F, I, and some S I ATP s.

Article Snippet: Samples were blocked with donkey serum for 1 h. Next, samples were incubated with rabbit anti-P2X1, mouse anti-P2X2, goat anti-P2X3, and chicken anti-P2X4 polyclonal antibodies (1∶200, Santa Cruz Biotechnology, Santa Cruz, CA) for 48 h. FITC- and TRITC- conjugated secondary antibodies were then incubated for 1 h.

Techniques: Patch Clamp, Microscopy, Immunohistochemistry, Negative Staining, Staining

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: Relevance of P2X1–4 subunits staining with four types of I ATP s.

Article Snippet: Samples were blocked with donkey serum for 1 h. Next, samples were incubated with rabbit anti-P2X1, mouse anti-P2X2, goat anti-P2X3, and chicken anti-P2X4 polyclonal antibodies (1∶200, Santa Cruz Biotechnology, Santa Cruz, CA) for 48 h. FITC- and TRITC- conjugated secondary antibodies were then incubated for 1 h.

Techniques: Staining

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: (A) A schematic diagram of the maximum cross section of a rat nodose ganglion (A, corresponding to panel B). (B) Representative graph of the distribution of P2X receptor-positive cells throughout the whole nodose ganglion section under a 20× light microscopic field. (C–F) Immunohistochemical staining using polyclonal antibodies against P2X1 (C), P2X2 (D), P2X3 (E), and P2X4 (F); 100× magnification. Scale bars in B = 500 µm and F = 100 µm.

Article Snippet: Sections were incubated with rabbit anti-P2X1, P2X2, P2X3, and P2X4 polyclonal antibodies (1∶1000, Cell Signaling Technology, USA) for 24 h at 4°C in a humid atmosphere.

Techniques: Immunohistochemical staining, Staining

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: (A), (B), (C), and (D) show P2X–IR sections that were counterstained with neutral red to precisely calculate the number of P2X1, P2X2, P2X3, and P2X4-positive neurons, respectively (using black positive cells compared with red negative cells). Scale bar in (D) = 100 µm. (E), (F), (G), and (H) represent the frequency distribution of the area-size of P2X1, P2X2, P2X3, and P2X4-IR, respectively.

Article Snippet: Sections were incubated with rabbit anti-P2X1, P2X2, P2X3, and P2X4 polyclonal antibodies (1∶1000, Cell Signaling Technology, USA) for 24 h at 4°C in a humid atmosphere.

Techniques:

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: The upper panel shows immunohistochemical staining for P2X1–4 subunits in the maximum cross section of the NG tissues. The lower panel shows that P2X3-IR was the most prevalent neuronal subtype, following by P2X2-IR; P2X4-IR had the lowest prevalence. * p <0.05.

Article Snippet: Sections were incubated with rabbit anti-P2X1, P2X2, P2X3, and P2X4 polyclonal antibodies (1∶1000, Cell Signaling Technology, USA) for 24 h at 4°C in a humid atmosphere.

Techniques: Immunohistochemical staining, Staining

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: NG neurons were stained using antibodies against P2X1, P2X2, P2X3, and P2X4 receptor subunits (A, D, G, and J, respectively). The nuclei of cultured NG neurons were stained with antibodies against NeuN (B, E, H, K). Merged images (C, F, I, L) representing co-staining of P2X receptor subunits and NeuN are shown. The scale bar shown in L is representative of all images, and represents 50 µm.

Article Snippet: Sections were incubated with rabbit anti-P2X1, P2X2, P2X3, and P2X4 polyclonal antibodies (1∶1000, Cell Signaling Technology, USA) for 24 h at 4°C in a humid atmosphere.

Techniques: Staining, Cell Culture

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: (A–C) Immunoreactivity of P2X1 and P2X2 (A–C), P2X1 and P2X3 (D–F), and P2X2 and P2X3 (G–I) subunits in the NG. Scale bars = 50 µm.

Article Snippet: Sections were incubated with rabbit anti-P2X1, P2X2, P2X3, and P2X4 polyclonal antibodies (1∶1000, Cell Signaling Technology, USA) for 24 h at 4°C in a humid atmosphere.

Techniques:

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: (A) Schematic view of the setup for the whole cell patch clamp and a representative image of a recorded cell under the phase contrast microscope and immunohistochemistry. (B) Immunohistochemistry revealed positive or negative staining for P2X1–4 subunits, which correlated with the type of I ATP and cell size. The samples in each row were from four different neurons that responded to ATP with different types of ATP-activated current. P2X3 staining was positive in all four types of I ATP neurons. P2X1 was positive in F, I, and S I ATP s, but negative in VS. P2X2 staining was only absent in neurons with type I I ATP , and P2X4 was positive in neurons with type F, I, and some S I ATP s.

Article Snippet: Sections were incubated with rabbit anti-P2X1, P2X2, P2X3, and P2X4 polyclonal antibodies (1∶1000, Cell Signaling Technology, USA) for 24 h at 4°C in a humid atmosphere.

Techniques: Patch Clamp, Microscopy, Immunohistochemistry, Negative Staining, Staining

Journal: PLoS ONE

Article Title: Comparative Analysis of P2X1, P2X2, P2X3, and P2X4 Receptor Subunits in Rat Nodose Ganglion Neurons

doi: 10.1371/journal.pone.0096699

Figure Lengend Snippet: Relevance of P2X1–4 subunits staining with four types of I ATP s.

Article Snippet: Sections were incubated with rabbit anti-P2X1, P2X2, P2X3, and P2X4 polyclonal antibodies (1∶1000, Cell Signaling Technology, USA) for 24 h at 4°C in a humid atmosphere.

Techniques: Staining