rabbit anti mouse p2x2 antibody  (Alomone Labs)


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    Alomone Labs rabbit anti mouse p2x2 antibody
    Compensatory mechanisms for the loss of P2x6 function in the heart. a-h) The mRNA expression levels of P2x1 (a), <t>P2x2</t> (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), Trpm7 (g), Cnnm2 (h), in heart of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data represent mean (n = 10) ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice. * P
    Rabbit Anti Mouse P2x2 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti mouse p2x2 antibody/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti mouse p2x2 antibody - by Bioz Stars, 2022-09
    86/100 stars

    Images

    1) Product Images from "P2X6 Knockout Mice Exhibit Normal Electrolyte Homeostasis"

    Article Title: P2X6 Knockout Mice Exhibit Normal Electrolyte Homeostasis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0156803

    Compensatory mechanisms for the loss of P2x6 function in the heart. a-h) The mRNA expression levels of P2x1 (a), P2x2 (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), Trpm7 (g), Cnnm2 (h), in heart of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data represent mean (n = 10) ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice. * P
    Figure Legend Snippet: Compensatory mechanisms for the loss of P2x6 function in the heart. a-h) The mRNA expression levels of P2x1 (a), P2x2 (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), Trpm7 (g), Cnnm2 (h), in heart of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data represent mean (n = 10) ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice. * P

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR

    Protein abundance of P2x2 and P2x4 in response to the loss of P2x6. a) Western blots of membrane and cytosol fractions of P2x6 -/- (KO) and P2x6 +/+ (WT) mice. The upper blot shows the immune-staining for P2x2 in mouse kidney material. To the right a western blot of HEK293 cells transiently transfected with HA-tagged P2x4 (P4) and mock (M) constructs is displayed. Middle, two western blots below are immune-stained for P2x4, left depicts P2x6 -/- (KO) and P2x6 +/+ (WT) material stained for P2x4, right represents a P2x4 blot on HEK293 material transiently transfected with human P2X4 and a mock construct. Bottom, displays a ß-actin immune-staining used as a loading control. Ladders (ez-run prestained marker (ThermoScientific, Breda, The Netherlands) represent protein size in kilo Dalton (kD). b) Protein expression levels for the P2x2 membrane lysate, P2x4 membrane lysate, P2x4 cytosol lysate, ß-actin membrane and ß-actin cytosol lysates in P2x6 -/- and P2x6 +/+ mice. Data (n = 3) represents mean ± SEM and are expressed as the % of total band intensity.
    Figure Legend Snippet: Protein abundance of P2x2 and P2x4 in response to the loss of P2x6. a) Western blots of membrane and cytosol fractions of P2x6 -/- (KO) and P2x6 +/+ (WT) mice. The upper blot shows the immune-staining for P2x2 in mouse kidney material. To the right a western blot of HEK293 cells transiently transfected with HA-tagged P2x4 (P4) and mock (M) constructs is displayed. Middle, two western blots below are immune-stained for P2x4, left depicts P2x6 -/- (KO) and P2x6 +/+ (WT) material stained for P2x4, right represents a P2x4 blot on HEK293 material transiently transfected with human P2X4 and a mock construct. Bottom, displays a ß-actin immune-staining used as a loading control. Ladders (ez-run prestained marker (ThermoScientific, Breda, The Netherlands) represent protein size in kilo Dalton (kD). b) Protein expression levels for the P2x2 membrane lysate, P2x4 membrane lysate, P2x4 cytosol lysate, ß-actin membrane and ß-actin cytosol lysates in P2x6 -/- and P2x6 +/+ mice. Data (n = 3) represents mean ± SEM and are expressed as the % of total band intensity.

    Techniques Used: Western Blot, Mouse Assay, Staining, Transfection, Construct, Marker, Expressing

    P2x subunit expression in response to the loss of P2x6 function in the kidney. a-f) The mRNA expression levels of P2x1 (a), P2x2 (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), in kidney of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data (n = 10) represent mean ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice.
    Figure Legend Snippet: P2x subunit expression in response to the loss of P2x6 function in the kidney. a-f) The mRNA expression levels of P2x1 (a), P2x2 (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), in kidney of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data (n = 10) represent mean ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice.

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR

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    Alomone Labs rabbit anti mouse p2x2 antibody
    Compensatory mechanisms for the loss of P2x6 function in the heart. a-h) The mRNA expression levels of P2x1 (a), <t>P2x2</t> (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), Trpm7 (g), Cnnm2 (h), in heart of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data represent mean (n = 10) ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice. * P
    Rabbit Anti Mouse P2x2 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti mouse p2x2 antibody/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti mouse p2x2 antibody - by Bioz Stars, 2022-09
    86/100 stars
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    96
    Alomone Labs rabbit anti p2x7 receptor
    Treatment with Andro reduces the upregulated co-expression values of the <t>P2X7</t> <t>receptor</t> and GFAP in the DRG of gp120+ddC-treated rats. (A) The co-expression of P2X7 receptor and GFAP was analyzed on the 18–20th day after surgery. Co-expression of the P2X7 receptor and GFAP in the gp120+ddC group was higher than that in the sham group. No difference was found between control+andro and sham rats. Treatment with Andro reduced the co-expression values of the P2X7 receptor and GFAP in gp120+ddC+andro-treated rats compared with gp120+ddC-treated rats that did not receive Andro treatment ( n = 8 for each group). Scale bar: 30 μm. (B) Histogram showed that the IOD value for co-expression of the P2X7 receptor and GFAP in the DRG. (C) Histogram showed that the number of neurons surrounded with GFAP- and P2X7-positive SGCs in the DRG SCGs. The data are shown as the mean ± SD. ∗∗ p
    Rabbit Anti P2x7 Receptor, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti p2x7 receptor/product/Alomone Labs
    Average 96 stars, based on 1 article reviews
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    94
    Alomone Labs rabbit anti p2x2
    Gustatory ganglion cells that innervate taste buds express Shh-tdTomato. (A) Shh CreERT2 ; R26R tdTomato (Shh-tdTomato) mice were given tamoxifen daily for 4 days and harvested at 10, 60 or 85 days. (B-D) In Shh-tdTomato + tongues, punctate signal (red) is evident in FFP at all times (arrowheads); at 60 and 85 days (C,D) gustatory innervation (arrows) associated with FFP (arrowheads) is also labeled. (E) Gustatory neurons in the geniculate ganglion (gVII) innervate FFP via the chorda tympani (CT) nerve, and project to the nucleus of the solitary tract (NST) in the brainstem. The greater superficial petrosal (GSP) nerve innervates soft palate taste buds (not shown). gVII neurons project centrally to the NST. (F-H) Cells in gVII express tdTomato at all time points. (I-K‴) At 60 days, PGP9.5 + nerve fibers (white in J,K″; green in K,K‴) innervate taste buds and adjacent FF epithelium, whereas Shh-tdTomato + neurites (white in I,K′; red in K,K‴), which are also PGP9.5 + , innervate taste buds exclusively (K′-K‴, cyan arrowheads). (L-N‴) <t>P2X2</t> + taste fibers (white in M,N′; green in N,N‴) express Shh-tdTomato (white in L,N′; red in N,N‴; cyan arrowheads). Boxes in K and N are shown at higher magnification in K′-K‴ and N′-N‴, respectively. Sparse bright lineage-labeled taste cells are detected at later times (I,L, arrows), and are distinguishable from dimmer Shh-tdTomato + neurites (I,L, arrowheads within taste buds). Nuclei are counterstained with Draq5 (blue). B-D and F-H are images of whole tongue and ganglia. I-N‴ are compressed confocal z -stacks. Scale bars: 1 mm in B-D; 150 μm in F-H; 10 μm in I-N‴.
    Rabbit Anti P2x2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti p2x2/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p2x2 - by Bioz Stars, 2022-09
    94/100 stars
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    Compensatory mechanisms for the loss of P2x6 function in the heart. a-h) The mRNA expression levels of P2x1 (a), P2x2 (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), Trpm7 (g), Cnnm2 (h), in heart of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data represent mean (n = 10) ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice. * P

    Journal: PLoS ONE

    Article Title: P2X6 Knockout Mice Exhibit Normal Electrolyte Homeostasis

    doi: 10.1371/journal.pone.0156803

    Figure Lengend Snippet: Compensatory mechanisms for the loss of P2x6 function in the heart. a-h) The mRNA expression levels of P2x1 (a), P2x2 (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), Trpm7 (g), Cnnm2 (h), in heart of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data represent mean (n = 10) ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice. * P

    Article Snippet: Samples were then subjected to SDS-PAGE and Western blots were incubated with a rabbit anti-mouse P2x2 antibody (Alomone Labs, Jerusalem, Israel, 1:500) or a rabbit anti-mouse P2x4 (H-40, Santa Cruz biotechnology, Santa Cruz CA, USA, 1:500) in 5% milk o/n at 4°C.

    Techniques: Expressing, Mouse Assay, Quantitative RT-PCR

    Protein abundance of P2x2 and P2x4 in response to the loss of P2x6. a) Western blots of membrane and cytosol fractions of P2x6 -/- (KO) and P2x6 +/+ (WT) mice. The upper blot shows the immune-staining for P2x2 in mouse kidney material. To the right a western blot of HEK293 cells transiently transfected with HA-tagged P2x4 (P4) and mock (M) constructs is displayed. Middle, two western blots below are immune-stained for P2x4, left depicts P2x6 -/- (KO) and P2x6 +/+ (WT) material stained for P2x4, right represents a P2x4 blot on HEK293 material transiently transfected with human P2X4 and a mock construct. Bottom, displays a ß-actin immune-staining used as a loading control. Ladders (ez-run prestained marker (ThermoScientific, Breda, The Netherlands) represent protein size in kilo Dalton (kD). b) Protein expression levels for the P2x2 membrane lysate, P2x4 membrane lysate, P2x4 cytosol lysate, ß-actin membrane and ß-actin cytosol lysates in P2x6 -/- and P2x6 +/+ mice. Data (n = 3) represents mean ± SEM and are expressed as the % of total band intensity.

    Journal: PLoS ONE

    Article Title: P2X6 Knockout Mice Exhibit Normal Electrolyte Homeostasis

    doi: 10.1371/journal.pone.0156803

    Figure Lengend Snippet: Protein abundance of P2x2 and P2x4 in response to the loss of P2x6. a) Western blots of membrane and cytosol fractions of P2x6 -/- (KO) and P2x6 +/+ (WT) mice. The upper blot shows the immune-staining for P2x2 in mouse kidney material. To the right a western blot of HEK293 cells transiently transfected with HA-tagged P2x4 (P4) and mock (M) constructs is displayed. Middle, two western blots below are immune-stained for P2x4, left depicts P2x6 -/- (KO) and P2x6 +/+ (WT) material stained for P2x4, right represents a P2x4 blot on HEK293 material transiently transfected with human P2X4 and a mock construct. Bottom, displays a ß-actin immune-staining used as a loading control. Ladders (ez-run prestained marker (ThermoScientific, Breda, The Netherlands) represent protein size in kilo Dalton (kD). b) Protein expression levels for the P2x2 membrane lysate, P2x4 membrane lysate, P2x4 cytosol lysate, ß-actin membrane and ß-actin cytosol lysates in P2x6 -/- and P2x6 +/+ mice. Data (n = 3) represents mean ± SEM and are expressed as the % of total band intensity.

    Article Snippet: Samples were then subjected to SDS-PAGE and Western blots were incubated with a rabbit anti-mouse P2x2 antibody (Alomone Labs, Jerusalem, Israel, 1:500) or a rabbit anti-mouse P2x4 (H-40, Santa Cruz biotechnology, Santa Cruz CA, USA, 1:500) in 5% milk o/n at 4°C.

    Techniques: Western Blot, Mouse Assay, Staining, Transfection, Construct, Marker, Expressing

    P2x subunit expression in response to the loss of P2x6 function in the kidney. a-f) The mRNA expression levels of P2x1 (a), P2x2 (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), in kidney of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data (n = 10) represent mean ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice.

    Journal: PLoS ONE

    Article Title: P2X6 Knockout Mice Exhibit Normal Electrolyte Homeostasis

    doi: 10.1371/journal.pone.0156803

    Figure Lengend Snippet: P2x subunit expression in response to the loss of P2x6 function in the kidney. a-f) The mRNA expression levels of P2x1 (a), P2x2 (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), in kidney of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data (n = 10) represent mean ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice.

    Article Snippet: Samples were then subjected to SDS-PAGE and Western blots were incubated with a rabbit anti-mouse P2x2 antibody (Alomone Labs, Jerusalem, Israel, 1:500) or a rabbit anti-mouse P2x4 (H-40, Santa Cruz biotechnology, Santa Cruz CA, USA, 1:500) in 5% milk o/n at 4°C.

    Techniques: Expressing, Mouse Assay, Quantitative RT-PCR

    Treatment with Andro reduces the upregulated co-expression values of the P2X7 receptor and GFAP in the DRG of gp120+ddC-treated rats. (A) The co-expression of P2X7 receptor and GFAP was analyzed on the 18–20th day after surgery. Co-expression of the P2X7 receptor and GFAP in the gp120+ddC group was higher than that in the sham group. No difference was found between control+andro and sham rats. Treatment with Andro reduced the co-expression values of the P2X7 receptor and GFAP in gp120+ddC+andro-treated rats compared with gp120+ddC-treated rats that did not receive Andro treatment ( n = 8 for each group). Scale bar: 30 μm. (B) Histogram showed that the IOD value for co-expression of the P2X7 receptor and GFAP in the DRG. (C) Histogram showed that the number of neurons surrounded with GFAP- and P2X7-positive SGCs in the DRG SCGs. The data are shown as the mean ± SD. ∗∗ p

    Journal: Frontiers in Pharmacology

    Article Title: Andrographolide Inhibits Mechanical and Thermal Hyperalgesia in a Rat Model of HIV-Induced Neuropathic Pain

    doi: 10.3389/fphar.2018.00593

    Figure Lengend Snippet: Treatment with Andro reduces the upregulated co-expression values of the P2X7 receptor and GFAP in the DRG of gp120+ddC-treated rats. (A) The co-expression of P2X7 receptor and GFAP was analyzed on the 18–20th day after surgery. Co-expression of the P2X7 receptor and GFAP in the gp120+ddC group was higher than that in the sham group. No difference was found between control+andro and sham rats. Treatment with Andro reduced the co-expression values of the P2X7 receptor and GFAP in gp120+ddC+andro-treated rats compared with gp120+ddC-treated rats that did not receive Andro treatment ( n = 8 for each group). Scale bar: 30 μm. (B) Histogram showed that the IOD value for co-expression of the P2X7 receptor and GFAP in the DRG. (C) Histogram showed that the number of neurons surrounded with GFAP- and P2X7-positive SGCs in the DRG SCGs. The data are shown as the mean ± SD. ∗∗ p

    Article Snippet: The membrane was blocked with 5% non-fat dry milk in 25 mM of Tris-buffered saline, pH 7.2, plus 0.05% Tween 20 (TBST) for 2 h at room temperature, followed by incubation with a rabbit anti-P2X7 receptor (1:200, Alomone Lab, Israel), or with other antibodies such as rabbit anti-tumor necrosis factor-α-receptor (TNF-α-R), rabbit anti-interleukin-1β (IL-1β) and mouse anti-IL-10 (1:500 dilutions, Abcam, Co.), anti-total extracellular regulated protein kinases (ERK)1/2 and anti-phospho-ERK1/2 MAPK (1:1000, Cell Signaling Technology), and mouse monoclonal anti-β-actin antibody (1:800 dilutions, Beijing Zhongshan Biotech Co., China) at 4°C overnight.

    Techniques: Expressing

    Molecular docking of rP2X7 protein and Andro. Simulation modeling of Andro docking with P2X7 protein was simulated by computer. (A) Front and aerial view showed that the best docking position between Andro and rP2X7. The docking position was in the outside of the cell membrane and at the waist of the protein structure, which is near the ATP docking pocket. (B) Showed that the best docking pocket between Andro and rP2X7. The ligand, Andro represented in yellow, and P2X7 protein’s active binding pocket displayed in a variety of colors. (C) Indicates a possible hydrogen bond formed between the connection residues (GLN143) and ligand. The data showed that Andro could interact with the P2X7 protein.

    Journal: Frontiers in Pharmacology

    Article Title: Andrographolide Inhibits Mechanical and Thermal Hyperalgesia in a Rat Model of HIV-Induced Neuropathic Pain

    doi: 10.3389/fphar.2018.00593

    Figure Lengend Snippet: Molecular docking of rP2X7 protein and Andro. Simulation modeling of Andro docking with P2X7 protein was simulated by computer. (A) Front and aerial view showed that the best docking position between Andro and rP2X7. The docking position was in the outside of the cell membrane and at the waist of the protein structure, which is near the ATP docking pocket. (B) Showed that the best docking pocket between Andro and rP2X7. The ligand, Andro represented in yellow, and P2X7 protein’s active binding pocket displayed in a variety of colors. (C) Indicates a possible hydrogen bond formed between the connection residues (GLN143) and ligand. The data showed that Andro could interact with the P2X7 protein.

    Article Snippet: The membrane was blocked with 5% non-fat dry milk in 25 mM of Tris-buffered saline, pH 7.2, plus 0.05% Tween 20 (TBST) for 2 h at room temperature, followed by incubation with a rabbit anti-P2X7 receptor (1:200, Alomone Lab, Israel), or with other antibodies such as rabbit anti-tumor necrosis factor-α-receptor (TNF-α-R), rabbit anti-interleukin-1β (IL-1β) and mouse anti-IL-10 (1:500 dilutions, Abcam, Co.), anti-total extracellular regulated protein kinases (ERK)1/2 and anti-phospho-ERK1/2 MAPK (1:1000, Cell Signaling Technology), and mouse monoclonal anti-β-actin antibody (1:800 dilutions, Beijing Zhongshan Biotech Co., China) at 4°C overnight.

    Techniques: Binding Assay

    Effects of the Andro treatment on the expression of P2X7 mRNA and protein in L4–6 DRGs from each group. (A) Expression of P2X7 mRNA in the DRG was measured by RT-qPCR ( n = 8 per group). Expression in the gp120+ddC group was higher than that in the sham group ( p

    Journal: Frontiers in Pharmacology

    Article Title: Andrographolide Inhibits Mechanical and Thermal Hyperalgesia in a Rat Model of HIV-Induced Neuropathic Pain

    doi: 10.3389/fphar.2018.00593

    Figure Lengend Snippet: Effects of the Andro treatment on the expression of P2X7 mRNA and protein in L4–6 DRGs from each group. (A) Expression of P2X7 mRNA in the DRG was measured by RT-qPCR ( n = 8 per group). Expression in the gp120+ddC group was higher than that in the sham group ( p

    Article Snippet: The membrane was blocked with 5% non-fat dry milk in 25 mM of Tris-buffered saline, pH 7.2, plus 0.05% Tween 20 (TBST) for 2 h at room temperature, followed by incubation with a rabbit anti-P2X7 receptor (1:200, Alomone Lab, Israel), or with other antibodies such as rabbit anti-tumor necrosis factor-α-receptor (TNF-α-R), rabbit anti-interleukin-1β (IL-1β) and mouse anti-IL-10 (1:500 dilutions, Abcam, Co.), anti-total extracellular regulated protein kinases (ERK)1/2 and anti-phospho-ERK1/2 MAPK (1:1000, Cell Signaling Technology), and mouse monoclonal anti-β-actin antibody (1:800 dilutions, Beijing Zhongshan Biotech Co., China) at 4°C overnight.

    Techniques: Expressing, Quantitative RT-PCR

    Gustatory ganglion cells that innervate taste buds express Shh-tdTomato. (A) Shh CreERT2 ; R26R tdTomato (Shh-tdTomato) mice were given tamoxifen daily for 4 days and harvested at 10, 60 or 85 days. (B-D) In Shh-tdTomato + tongues, punctate signal (red) is evident in FFP at all times (arrowheads); at 60 and 85 days (C,D) gustatory innervation (arrows) associated with FFP (arrowheads) is also labeled. (E) Gustatory neurons in the geniculate ganglion (gVII) innervate FFP via the chorda tympani (CT) nerve, and project to the nucleus of the solitary tract (NST) in the brainstem. The greater superficial petrosal (GSP) nerve innervates soft palate taste buds (not shown). gVII neurons project centrally to the NST. (F-H) Cells in gVII express tdTomato at all time points. (I-K‴) At 60 days, PGP9.5 + nerve fibers (white in J,K″; green in K,K‴) innervate taste buds and adjacent FF epithelium, whereas Shh-tdTomato + neurites (white in I,K′; red in K,K‴), which are also PGP9.5 + , innervate taste buds exclusively (K′-K‴, cyan arrowheads). (L-N‴) P2X2 + taste fibers (white in M,N′; green in N,N‴) express Shh-tdTomato (white in L,N′; red in N,N‴; cyan arrowheads). Boxes in K and N are shown at higher magnification in K′-K‴ and N′-N‴, respectively. Sparse bright lineage-labeled taste cells are detected at later times (I,L, arrows), and are distinguishable from dimmer Shh-tdTomato + neurites (I,L, arrowheads within taste buds). Nuclei are counterstained with Draq5 (blue). B-D and F-H are images of whole tongue and ganglia. I-N‴ are compressed confocal z -stacks. Scale bars: 1 mm in B-D; 150 μm in F-H; 10 μm in I-N‴.

    Journal: Development (Cambridge, England)

    Article Title: Sonic hedgehog from both nerves and epithelium is a key trophic factor for taste bud maintenance

    doi: 10.1242/dev.150342

    Figure Lengend Snippet: Gustatory ganglion cells that innervate taste buds express Shh-tdTomato. (A) Shh CreERT2 ; R26R tdTomato (Shh-tdTomato) mice were given tamoxifen daily for 4 days and harvested at 10, 60 or 85 days. (B-D) In Shh-tdTomato + tongues, punctate signal (red) is evident in FFP at all times (arrowheads); at 60 and 85 days (C,D) gustatory innervation (arrows) associated with FFP (arrowheads) is also labeled. (E) Gustatory neurons in the geniculate ganglion (gVII) innervate FFP via the chorda tympani (CT) nerve, and project to the nucleus of the solitary tract (NST) in the brainstem. The greater superficial petrosal (GSP) nerve innervates soft palate taste buds (not shown). gVII neurons project centrally to the NST. (F-H) Cells in gVII express tdTomato at all time points. (I-K‴) At 60 days, PGP9.5 + nerve fibers (white in J,K″; green in K,K‴) innervate taste buds and adjacent FF epithelium, whereas Shh-tdTomato + neurites (white in I,K′; red in K,K‴), which are also PGP9.5 + , innervate taste buds exclusively (K′-K‴, cyan arrowheads). (L-N‴) P2X2 + taste fibers (white in M,N′; green in N,N‴) express Shh-tdTomato (white in L,N′; red in N,N‴; cyan arrowheads). Boxes in K and N are shown at higher magnification in K′-K‴ and N′-N‴, respectively. Sparse bright lineage-labeled taste cells are detected at later times (I,L, arrows), and are distinguishable from dimmer Shh-tdTomato + neurites (I,L, arrowheads within taste buds). Nuclei are counterstained with Draq5 (blue). B-D and F-H are images of whole tongue and ganglia. I-N‴ are compressed confocal z -stacks. Scale bars: 1 mm in B-D; 150 μm in F-H; 10 μm in I-N‴.

    Article Snippet: Primary antibodies used were: rat anti-K8 (Troma) (1:250; DSHB, University of Iowa; RRID AB_531826), rabbit anti-P2X2 (1:500; APR-003, Alomone Labs; RRID:AB_2040054), rabbit anti-PGP9.5 (1:1000; 7863-0504, AbD Serotec; RRID AB_2210505), rabbit anti-Ki67 (1:200; RM-9106-S, Thermo Fisher Scientific; RRID:AB_2341197) and chicken anti-GFP to detect YFP (1:1000; GFP-1020, Aves Labs; RRID:AB_10000240).

    Techniques: Mouse Assay, Labeling

    Genetic deletion of Shh in Shh + cells, including gustatory neurons, minimally affects taste buds. (A) Shh CreERT2/flox ; R26R tdTomato (Shh-ShhcKO) and genetic control mice were given tamoxifen for 4 days and harvested at 35 days. (B) In tamoxifen-treated Shh-ShhcKO mice, Shh is deleted permanently from ganglion cells and nerves (indicated by the cross), but transiently from taste buds (see text). (C,D) tdTomato reports Shh deletion in tongue (C) and gVII (D) of mutant mice. (E) Quantitative PCR reveals that neither Shh nor Gli1 expression is significantly reduced in mutant tongue epithelium. (F) Expression of Shh , but not Gli1, is significantly reduced in mutant gVII. (G,H) Typical FFP number does not differ between mutants and controls (G), although atypical FFP increase in mutants (H). (I,J) The size of taste buds in typical (I) and atypical (J) FFP in mutants does not differ from controls. (K) Shh-tdTomato + neurites (red) innervate a taste bud (K8 + , green) in an atypical FFP in a Shh-ShhcKO mouse (Shh-descendent taste cell, arrow). (L) The proportion of FFP innervated by P2X2 + fibers is not affected by Shh-ShhcKO. (M,N) P2X2 + innervation density of taste buds in typical (M) and atypical (N) FFP does not differ between controls and Shh-ShhcKO mice. Nuclei counterstained with Draq5 (blue); white dashed lines delimit basement membrane; white solid lines delimit epithelial surface. C and D are images of whole tongue and gVII. K is a compressed confocal z -stack. Scale bars: 1 mm in C; 150 μm in D; 10 μm in K. n =3-5 mice per condition. Data are mean±s.d., except E and F, which are mean±s.e.m; I and N represent the median with interquartile range. Results were analyzed using Student's t -test (E-H,J,L,M) or Mann–Whitney U-test (I,N). *** P

    Journal: Development (Cambridge, England)

    Article Title: Sonic hedgehog from both nerves and epithelium is a key trophic factor for taste bud maintenance

    doi: 10.1242/dev.150342

    Figure Lengend Snippet: Genetic deletion of Shh in Shh + cells, including gustatory neurons, minimally affects taste buds. (A) Shh CreERT2/flox ; R26R tdTomato (Shh-ShhcKO) and genetic control mice were given tamoxifen for 4 days and harvested at 35 days. (B) In tamoxifen-treated Shh-ShhcKO mice, Shh is deleted permanently from ganglion cells and nerves (indicated by the cross), but transiently from taste buds (see text). (C,D) tdTomato reports Shh deletion in tongue (C) and gVII (D) of mutant mice. (E) Quantitative PCR reveals that neither Shh nor Gli1 expression is significantly reduced in mutant tongue epithelium. (F) Expression of Shh , but not Gli1, is significantly reduced in mutant gVII. (G,H) Typical FFP number does not differ between mutants and controls (G), although atypical FFP increase in mutants (H). (I,J) The size of taste buds in typical (I) and atypical (J) FFP in mutants does not differ from controls. (K) Shh-tdTomato + neurites (red) innervate a taste bud (K8 + , green) in an atypical FFP in a Shh-ShhcKO mouse (Shh-descendent taste cell, arrow). (L) The proportion of FFP innervated by P2X2 + fibers is not affected by Shh-ShhcKO. (M,N) P2X2 + innervation density of taste buds in typical (M) and atypical (N) FFP does not differ between controls and Shh-ShhcKO mice. Nuclei counterstained with Draq5 (blue); white dashed lines delimit basement membrane; white solid lines delimit epithelial surface. C and D are images of whole tongue and gVII. K is a compressed confocal z -stack. Scale bars: 1 mm in C; 150 μm in D; 10 μm in K. n =3-5 mice per condition. Data are mean±s.d., except E and F, which are mean±s.e.m; I and N represent the median with interquartile range. Results were analyzed using Student's t -test (E-H,J,L,M) or Mann–Whitney U-test (I,N). *** P

    Article Snippet: Primary antibodies used were: rat anti-K8 (Troma) (1:250; DSHB, University of Iowa; RRID AB_531826), rabbit anti-P2X2 (1:500; APR-003, Alomone Labs; RRID:AB_2040054), rabbit anti-PGP9.5 (1:1000; 7863-0504, AbD Serotec; RRID AB_2210505), rabbit anti-Ki67 (1:200; RM-9106-S, Thermo Fisher Scientific; RRID:AB_2341197) and chicken anti-GFP to detect YFP (1:1000; GFP-1020, Aves Labs; RRID:AB_10000240).

    Techniques: Mouse Assay, Mutagenesis, Real-time Polymerase Chain Reaction, Expressing, MANN-WHITNEY