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  • 93
    Sino Biological vglut2 plasmid cdna
    Unaltered DA neuronal development in the absence of <t>VGluT2,</t> but regional striatal innervation deficit. A , Immunohistochemistry analysis against TH and DAPI in coronal slices of E18.5 VGluT2WT and constitutive VGluT2KO littermates reveals no macroscopic changes in the mesencephalic dopaminergic system. Scale bar, 100 µm. B , Quantification of the number of TH + neurons at E18.5 in the mesencephalon reveals no significant differences between VGluT2WT and VGluT2KO littermates. C , Immunohistochemistry analysis against TH and DAPI in coronal slices of E18.5 VGluT2WT and VGluT2KO littermates reveals no macroscopic changes in the striatum. Scale bar, 100 µm. D , E , Quantification of TH-immunoreactivity in the striatum at E18.5 reveals a regional decrease in intensity in caudal, dorsal striatum ( D : n = 4 pups, p
    Vglut2 Plasmid Cdna, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vglut2 plasmid cdna/product/Sino Biological
    Average 93 stars, based on 1 article reviews
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    93
    Sino Biological ankrd11
    The variants of <t>ANKRD11</t> attenuate p21 . (a) Knockdown efficiency of shRNAs for ANKRD11 by immunoblotting. HEK293T cells were transfected with shRNAs (scramble, sh1 or sh2) for ANKRD11, and the protein levels of ANKRD11 and GAPDH were detected by immunoblotting. Sh1 and sh2 reduced the expression of ANKRD11 protein, respectively. (b) Luciferase reporter assay to detect the relative activities of human p21‐ promoter. HEK293T cells were transfected with shRNAs for ANKRD11, as well as with p21‐ promoter luciferase reporter and PRL‐TK. The cell lysates subjected to Dual‐Luciferase Reporter assay 48 hrs later. Data are expressed as mean ± SD. ** p
    Ankrd11, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ankrd11/product/Sino Biological
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    97
    Millipore lipopolysaccharide
    The variants of <t>ANKRD11</t> attenuate p21 . (a) Knockdown efficiency of shRNAs for ANKRD11 by immunoblotting. HEK293T cells were transfected with shRNAs (scramble, sh1 or sh2) for ANKRD11, and the protein levels of ANKRD11 and GAPDH were detected by immunoblotting. Sh1 and sh2 reduced the expression of ANKRD11 protein, respectively. (b) Luciferase reporter assay to detect the relative activities of human p21‐ promoter. HEK293T cells were transfected with shRNAs for ANKRD11, as well as with p21‐ promoter luciferase reporter and PRL‐TK. The cell lysates subjected to Dual‐Luciferase Reporter assay 48 hrs later. Data are expressed as mean ± SD. ** p
    Lipopolysaccharide, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Cayman Chemical phorbol 12 myristate 13 acetate
    The variants of <t>ANKRD11</t> attenuate p21 . (a) Knockdown efficiency of shRNAs for ANKRD11 by immunoblotting. HEK293T cells were transfected with shRNAs (scramble, sh1 or sh2) for ANKRD11, and the protein levels of ANKRD11 and GAPDH were detected by immunoblotting. Sh1 and sh2 reduced the expression of ANKRD11 protein, respectively. (b) Luciferase reporter assay to detect the relative activities of human p21‐ promoter. HEK293T cells were transfected with shRNAs for ANKRD11, as well as with p21‐ promoter luciferase reporter and PRL‐TK. The cell lysates subjected to Dual‐Luciferase Reporter assay 48 hrs later. Data are expressed as mean ± SD. ** p
    Phorbol 12 Myristate 13 Acetate, supplied by Cayman Chemical, 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/phorbol 12 myristate 13 acetate/product/Cayman Chemical
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    Image Search Results


    Unaltered DA neuronal development in the absence of VGluT2, but regional striatal innervation deficit. A , Immunohistochemistry analysis against TH and DAPI in coronal slices of E18.5 VGluT2WT and constitutive VGluT2KO littermates reveals no macroscopic changes in the mesencephalic dopaminergic system. Scale bar, 100 µm. B , Quantification of the number of TH + neurons at E18.5 in the mesencephalon reveals no significant differences between VGluT2WT and VGluT2KO littermates. C , Immunohistochemistry analysis against TH and DAPI in coronal slices of E18.5 VGluT2WT and VGluT2KO littermates reveals no macroscopic changes in the striatum. Scale bar, 100 µm. D , E , Quantification of TH-immunoreactivity in the striatum at E18.5 reveals a regional decrease in intensity in caudal, dorsal striatum ( D : n = 4 pups, p

    Journal: The Journal of Neuroscience

    Article Title: VGluT2 Expression in Dopamine Neurons Contributes to Postlesional Striatal Reinnervation

    doi: 10.1523/JNEUROSCI.0823-20.2020

    Figure Lengend Snippet: Unaltered DA neuronal development in the absence of VGluT2, but regional striatal innervation deficit. A , Immunohistochemistry analysis against TH and DAPI in coronal slices of E18.5 VGluT2WT and constitutive VGluT2KO littermates reveals no macroscopic changes in the mesencephalic dopaminergic system. Scale bar, 100 µm. B , Quantification of the number of TH + neurons at E18.5 in the mesencephalon reveals no significant differences between VGluT2WT and VGluT2KO littermates. C , Immunohistochemistry analysis against TH and DAPI in coronal slices of E18.5 VGluT2WT and VGluT2KO littermates reveals no macroscopic changes in the striatum. Scale bar, 100 µm. D , E , Quantification of TH-immunoreactivity in the striatum at E18.5 reveals a regional decrease in intensity in caudal, dorsal striatum ( D : n = 4 pups, p

    Article Snippet: Calculation of the absolute copy number for Th and VgluT2 in each cell was determined with an external calibration curve based on a Th or VGluT2 plasmid cDNA (obtained, respectively, from Sino Biological and Harvard University, Cambridge, MA).

    Techniques: Immunohistochemistry

    Most DA neurons have a Vglut2 expression history. A , Schematic representation of the intersectional genetic approach used the following: mice expressing a Th-Flpo and VGluT2-Cre conditional construct will express tdTomato only in cells that have expressed both Th and Vglut2 genes. B , C , A total of 98% of SN and VTA TH + neurons colocalize with tdTomato. Only a small number of TH + neurons are negative for tdTomato (yellow arrowhead). In the medial midbrain, tdTomato + neurons are found that are no longer positive for TH (yellow arrow). Scale bar, 200 µm. D , Brains taken from P1 VGluT2Cre + ;ThFlp-;tdTomato or VGluT2Cre-;ThFlp + ;tdTomato pups showed no tdTomato expression. Scale bar, 200 µm. E , Fluorescence in situ hybridization reveals that Vglut2 expression overlaps with that of Th in the mesencephalon at E11.5. Scale bar, 100 µm. F , TH-GFP + neurons taken at E11.5 from the mesencephalon express abundant VGluT2 protein after 24 h in culture (DIV1), both in the soma and growth cone (white box). Scale bar, 10 µm. G , Vglut2 transcript is still present in the mesencephalon at E14.5 and only partly overlaps with Th transcript in medial sections of the mesencephalon, but not in lateral sections. Scale bar, 25 µm.

    Journal: The Journal of Neuroscience

    Article Title: VGluT2 Expression in Dopamine Neurons Contributes to Postlesional Striatal Reinnervation

    doi: 10.1523/JNEUROSCI.0823-20.2020

    Figure Lengend Snippet: Most DA neurons have a Vglut2 expression history. A , Schematic representation of the intersectional genetic approach used the following: mice expressing a Th-Flpo and VGluT2-Cre conditional construct will express tdTomato only in cells that have expressed both Th and Vglut2 genes. B , C , A total of 98% of SN and VTA TH + neurons colocalize with tdTomato. Only a small number of TH + neurons are negative for tdTomato (yellow arrowhead). In the medial midbrain, tdTomato + neurons are found that are no longer positive for TH (yellow arrow). Scale bar, 200 µm. D , Brains taken from P1 VGluT2Cre + ;ThFlp-;tdTomato or VGluT2Cre-;ThFlp + ;tdTomato pups showed no tdTomato expression. Scale bar, 200 µm. E , Fluorescence in situ hybridization reveals that Vglut2 expression overlaps with that of Th in the mesencephalon at E11.5. Scale bar, 100 µm. F , TH-GFP + neurons taken at E11.5 from the mesencephalon express abundant VGluT2 protein after 24 h in culture (DIV1), both in the soma and growth cone (white box). Scale bar, 10 µm. G , Vglut2 transcript is still present in the mesencephalon at E14.5 and only partly overlaps with Th transcript in medial sections of the mesencephalon, but not in lateral sections. Scale bar, 25 µm.

    Article Snippet: Calculation of the absolute copy number for Th and VgluT2 in each cell was determined with an external calibration curve based on a Th or VGluT2 plasmid cDNA (obtained, respectively, from Sino Biological and Harvard University, Cambridge, MA).

    Techniques: Expressing, Mouse Assay, Construct, Fluorescence, In Situ Hybridization

    Proposed model of Vglut2 expression postlesional plasticity in DA neurons. A , Schematic diagram of a model for the plasticity of VGluT2 expression in DA neurons. Vglut2 is expressed in all DA neurons during early embryonic development, after which it is downregulated during maturation, at which point it is expressed in only a minority of DA neurons. Based on our single-cell qPCR experiments, we observed that ∼50% of DA neurons maintain ≥10 copies of Vglut2 (green line). After a lesion, ∼50% of SN DA neurons may have the capacity to reactivate Vglut2 expression above threshold (green line), while the other half of SN DA neurons continue to repress Vglut2 expression (red line). B , In vitro studies revealed that VGluT2 is expressed in growth cones of primary DA neurons, which allows the release of glutamate, which in turn can stimulate glutamate receptors on DA neurons and promote axonal outgrowth. Overexpression of VGluT2 in DA neurons results in enhanced axonal arborization, which could potentially be because of enhanced glutamate release by DA neurons. C , Here we propose that enhanced VGluT2 expression by DA neurons in the postlesional brain could reactivate the autocrine release of glutamate that can contribute to the axonal outgrowth of DA neurons toward target cells in the striatum.

    Journal: The Journal of Neuroscience

    Article Title: VGluT2 Expression in Dopamine Neurons Contributes to Postlesional Striatal Reinnervation

    doi: 10.1523/JNEUROSCI.0823-20.2020

    Figure Lengend Snippet: Proposed model of Vglut2 expression postlesional plasticity in DA neurons. A , Schematic diagram of a model for the plasticity of VGluT2 expression in DA neurons. Vglut2 is expressed in all DA neurons during early embryonic development, after which it is downregulated during maturation, at which point it is expressed in only a minority of DA neurons. Based on our single-cell qPCR experiments, we observed that ∼50% of DA neurons maintain ≥10 copies of Vglut2 (green line). After a lesion, ∼50% of SN DA neurons may have the capacity to reactivate Vglut2 expression above threshold (green line), while the other half of SN DA neurons continue to repress Vglut2 expression (red line). B , In vitro studies revealed that VGluT2 is expressed in growth cones of primary DA neurons, which allows the release of glutamate, which in turn can stimulate glutamate receptors on DA neurons and promote axonal outgrowth. Overexpression of VGluT2 in DA neurons results in enhanced axonal arborization, which could potentially be because of enhanced glutamate release by DA neurons. C , Here we propose that enhanced VGluT2 expression by DA neurons in the postlesional brain could reactivate the autocrine release of glutamate that can contribute to the axonal outgrowth of DA neurons toward target cells in the striatum.

    Article Snippet: Calculation of the absolute copy number for Th and VgluT2 in each cell was determined with an external calibration curve based on a Th or VGluT2 plasmid cDNA (obtained, respectively, from Sino Biological and Harvard University, Cambridge, MA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, In Vitro, Over Expression

    The variants of ANKRD11 attenuate p21 . (a) Knockdown efficiency of shRNAs for ANKRD11 by immunoblotting. HEK293T cells were transfected with shRNAs (scramble, sh1 or sh2) for ANKRD11, and the protein levels of ANKRD11 and GAPDH were detected by immunoblotting. Sh1 and sh2 reduced the expression of ANKRD11 protein, respectively. (b) Luciferase reporter assay to detect the relative activities of human p21‐ promoter. HEK293T cells were transfected with shRNAs for ANKRD11, as well as with p21‐ promoter luciferase reporter and PRL‐TK. The cell lysates subjected to Dual‐Luciferase Reporter assay 48 hrs later. Data are expressed as mean ± SD. ** p

    Journal: American Journal of Medical Genetics. Part a

    Article Title: Two loss‐of‐function ANKRD11 variants in Chinese patients with short stature and a possible molecular pathway, et al. Two loss‐of‐function ANKRD11 variants in Chinese patients with short stature and a possible molecular pathway

    doi: 10.1002/ajmg.a.62024

    Figure Lengend Snippet: The variants of ANKRD11 attenuate p21 . (a) Knockdown efficiency of shRNAs for ANKRD11 by immunoblotting. HEK293T cells were transfected with shRNAs (scramble, sh1 or sh2) for ANKRD11, and the protein levels of ANKRD11 and GAPDH were detected by immunoblotting. Sh1 and sh2 reduced the expression of ANKRD11 protein, respectively. (b) Luciferase reporter assay to detect the relative activities of human p21‐ promoter. HEK293T cells were transfected with shRNAs for ANKRD11, as well as with p21‐ promoter luciferase reporter and PRL‐TK. The cell lysates subjected to Dual‐Luciferase Reporter assay 48 hrs later. Data are expressed as mean ± SD. ** p

    Article Snippet: 2.3 Plasmids construction The cDNA containing ANKRD11 , which was purchased from Sino Biological (HG20681‐UT, Beijing, China), was used as template, and inserted into pCDNA3.0 plasmids with 5'flag tag.

    Techniques: Transfection, Expressing, Luciferase, Reporter Assay

    Clinical characteristics of two probands with ANKRD11 variants. (a‐1) Proband 1 had a triangular face, large cheek bone, low bridge of the nose and normal teeth. (a‐2) X‐ray examination of proband 1 at the age of 8 years and 4 months showing a delayed bone age (6.83 years) (by the Greulich‐Pyle method). (a‐3) X‐ray examination of proband 1 at the age of 10 years and 8 months showing an advanced bone age (12 years) (by the Greulich‐Pyle method). (a‐4) Growth curves of proband 1 showed an unideal growth tendency. Green arrow: delayed bone age; Blue arrow: advanced bone age. (b‐1) X‐ray examination showed an advanced bone age of proband 2 (by the Greulich‐Pyle method). (b‐2) Growth curves of proband 2 showed an unideal growth tendency. Approximate growth tendency from 6 years old to 9 years old was indicated by dotted line. Blue arrow: advanced bone age [Color figure can be viewed at wileyonlinelibrary.com ]

    Journal: American Journal of Medical Genetics. Part a

    Article Title: Two loss‐of‐function ANKRD11 variants in Chinese patients with short stature and a possible molecular pathway, et al. Two loss‐of‐function ANKRD11 variants in Chinese patients with short stature and a possible molecular pathway

    doi: 10.1002/ajmg.a.62024

    Figure Lengend Snippet: Clinical characteristics of two probands with ANKRD11 variants. (a‐1) Proband 1 had a triangular face, large cheek bone, low bridge of the nose and normal teeth. (a‐2) X‐ray examination of proband 1 at the age of 8 years and 4 months showing a delayed bone age (6.83 years) (by the Greulich‐Pyle method). (a‐3) X‐ray examination of proband 1 at the age of 10 years and 8 months showing an advanced bone age (12 years) (by the Greulich‐Pyle method). (a‐4) Growth curves of proband 1 showed an unideal growth tendency. Green arrow: delayed bone age; Blue arrow: advanced bone age. (b‐1) X‐ray examination showed an advanced bone age of proband 2 (by the Greulich‐Pyle method). (b‐2) Growth curves of proband 2 showed an unideal growth tendency. Approximate growth tendency from 6 years old to 9 years old was indicated by dotted line. Blue arrow: advanced bone age [Color figure can be viewed at wileyonlinelibrary.com ]

    Article Snippet: 2.3 Plasmids construction The cDNA containing ANKRD11 , which was purchased from Sino Biological (HG20681‐UT, Beijing, China), was used as template, and inserted into pCDNA3.0 plasmids with 5'flag tag.

    Techniques:

    The variants of ANKRD11 have no effect on the expression of its protein and mRNA. (a) Schematic view of human ANKRD11 protein showing the location of the two variants involved in our study. ANK, Ankyrin repeats; RD1, Repression domain 1; AD, Activation domain; RD2, Repression domain 2. (b) Detection of the relative mRNA levels of wild type ANKRD11 and its mutants by RT‐qPCR. Empty vector, wild type ANKRD11, ANKRD11(p. Lys1347del) or ANKRD11 (p. Leu2143Val) were transfected into HEK293T cells. Total RNA was extracted, complementary DNA (cDNA) was synthesized, and then the relative mRNA levels of ANKRND11 were detected by RT‐qPCR. Data are expressed as mean ± SD. ** p

    Journal: American Journal of Medical Genetics. Part a

    Article Title: Two loss‐of‐function ANKRD11 variants in Chinese patients with short stature and a possible molecular pathway, et al. Two loss‐of‐function ANKRD11 variants in Chinese patients with short stature and a possible molecular pathway

    doi: 10.1002/ajmg.a.62024

    Figure Lengend Snippet: The variants of ANKRD11 have no effect on the expression of its protein and mRNA. (a) Schematic view of human ANKRD11 protein showing the location of the two variants involved in our study. ANK, Ankyrin repeats; RD1, Repression domain 1; AD, Activation domain; RD2, Repression domain 2. (b) Detection of the relative mRNA levels of wild type ANKRD11 and its mutants by RT‐qPCR. Empty vector, wild type ANKRD11, ANKRD11(p. Lys1347del) or ANKRD11 (p. Leu2143Val) were transfected into HEK293T cells. Total RNA was extracted, complementary DNA (cDNA) was synthesized, and then the relative mRNA levels of ANKRND11 were detected by RT‐qPCR. Data are expressed as mean ± SD. ** p

    Article Snippet: 2.3 Plasmids construction The cDNA containing ANKRD11 , which was purchased from Sino Biological (HG20681‐UT, Beijing, China), was used as template, and inserted into pCDNA3.0 plasmids with 5'flag tag.

    Techniques: Expressing, Activation Assay, Quantitative RT-PCR, Plasmid Preparation, Transfection, Synthesized

    The pedigrees of two families with ANKRD11 variants. (a) Pedigree of the family of proband 1 with the c.4039_4041del mutation in ANKRD11 gene, and partial sequencing chromatographs of the proband and the parents. (b) Pedigree of the family of proband 2 with a c.6427C > G mutation in ANKRD11 gene, and partial sequencing chromatographs of the proband and the parents. Squares and circles in gray indicate family members who had not received mutation analysis [Color figure can be viewed at wileyonlinelibrary.com ]

    Journal: American Journal of Medical Genetics. Part a

    Article Title: Two loss‐of‐function ANKRD11 variants in Chinese patients with short stature and a possible molecular pathway, et al. Two loss‐of‐function ANKRD11 variants in Chinese patients with short stature and a possible molecular pathway

    doi: 10.1002/ajmg.a.62024

    Figure Lengend Snippet: The pedigrees of two families with ANKRD11 variants. (a) Pedigree of the family of proband 1 with the c.4039_4041del mutation in ANKRD11 gene, and partial sequencing chromatographs of the proband and the parents. (b) Pedigree of the family of proband 2 with a c.6427C > G mutation in ANKRD11 gene, and partial sequencing chromatographs of the proband and the parents. Squares and circles in gray indicate family members who had not received mutation analysis [Color figure can be viewed at wileyonlinelibrary.com ]

    Article Snippet: 2.3 Plasmids construction The cDNA containing ANKRD11 , which was purchased from Sino Biological (HG20681‐UT, Beijing, China), was used as template, and inserted into pCDNA3.0 plasmids with 5'flag tag.

    Techniques: Mutagenesis, Sequencing