myc p53 Search Results


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  • 99
    Santa Cruz Biotechnology p53
    Ribosomal stress leads to RPS2 ubiquitination for MDM2 regulation to induce <t>p53</t> by dissociation of USP47 and RPS2. ( a ) U2OS cells co-overexpressing either Flag vector or Flag-USP47 were treated with DMSO or 5 nM actinomycin D for 4 h, and immunoprecipitation was performed with anti-Flag agarose. Bound proteins were immunoblotted with the indicated antibodies. α-tubulin was used as a loading control. ( b ) HEK293T cells were co-overexpressed with HA-RPS2, Flag-USP47, and then treated with DMSO or 5 nM actinomycin D for 4 h. Then, cell lysates were immunoprecipitated with anti-Flag M2 agarose and detected with the indicated antibodies. ( c ) U2OS cells were co-overexpressed with Flag-USP47 and HA-RPS2 and then treated with DMSO or 5 nM actinomycin D for 4 h. Cells were immunostained with anti-HA antibody followed by Alexa Fluor 488 anti-mouse IgG and Flag antibodies followed by Alexa Fluor 546 anti-rabbit IgG. Representative immunofluorescence images were captured by confocal microscopy. ( d ) HEK293T cells were overexpressed with HA-RPS2 and His-ubiquitin and then treated with 10 μM MG132 for 4 h and 5 nM actinomycin D for indicated times. Ubiquitin conjugates were purified on Ni-NTA-agarose under denaturing conditions, and ubiquitinated RPS2 was detected by anti-HA antibodies. ( e ) U2OS cells were co-transfected with His-ubiquitin, HA-RPS2, and either siControl or siUSP47 and then treated with 10 μM MG132 and either DMSO or 5 nM actinomycin D for 4 h. Ubiquitin conjugates were purified on Ni-NTA agarose under denaturing conditions, and ubiquitinated RPS2 was detected by anti-HA antibodies. ( f ) U2OS cells transfected with siRNA against control or USP47 were treated with either DMSO or 5 nM actinomycin D for 4 h. Cell lysates were immunoblotted with the indicated antibodies. The uncropped blots and molecular weight markers are shown in Figure S7 .
    P53, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    OriGene human p53
    Inhibition of <t>p53</t> transactivation function overcomes zinc deficiency-induced failure of neural tube closure. (A) Still frames from time-lapse live imaging of E8.5 Ven Myr embryos show that an inhibitor of p53 transactivation function, pifithrin-α, but not an inhibitor of p53 mitochondrial function, pifithrin-µ, can rescue TPEN-induced failure of neural tube closure. (B) Time to neural tube closure after TPEN+pifithrin-α ( n =6 from three independent experiments) relative to control ( n =6 from four independent experiments) (the neural tube never closes with TPEN treatment) ( n =6 from two independent experiments) or TPEN +pifithrin-µ-treated embryos ( n =5 from two independent experiments). (C-F) Representative still images (C) and quantitative data (D-F) of time-lapse imaging show the ability of pifithrin-α to rescue TPEN-induced failure of neural tube closure. In A and C, the white lines, red lines, yellow lines and purple line show the edges of neural fold. In D, the pink arrowhead indicates failure of neural tube closure; the dark gray and green arrowheads indicate the timepoint of neural tube closure. * P
    Human P53, supplied by OriGene, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    p53  (Roche)
    97
    Roche p53
    Representative images of Immunohistochemistry for molecular markers of glioblastoma. Images for Ki67 (high above and low below), <t>p53</t> (positive staining above and negative below), and IDH1 (positive staining above and negative below) are presented.
    P53, supplied by Roche, 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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    97
    Thermo Fisher hct116
    Representative images of Immunohistochemistry for molecular markers of glioblastoma. Images for Ki67 (high above and low below), <t>p53</t> (positive staining above and negative below), and IDH1 (positive staining above and negative below) are presented.
    Hct116, supplied by Thermo Fisher, 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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    Full length Clone DNA of Human tumor protein p53 TP53 transcript variant 1 with C terminal Myc tag
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    Image Search Results


    Ribosomal stress leads to RPS2 ubiquitination for MDM2 regulation to induce p53 by dissociation of USP47 and RPS2. ( a ) U2OS cells co-overexpressing either Flag vector or Flag-USP47 were treated with DMSO or 5 nM actinomycin D for 4 h, and immunoprecipitation was performed with anti-Flag agarose. Bound proteins were immunoblotted with the indicated antibodies. α-tubulin was used as a loading control. ( b ) HEK293T cells were co-overexpressed with HA-RPS2, Flag-USP47, and then treated with DMSO or 5 nM actinomycin D for 4 h. Then, cell lysates were immunoprecipitated with anti-Flag M2 agarose and detected with the indicated antibodies. ( c ) U2OS cells were co-overexpressed with Flag-USP47 and HA-RPS2 and then treated with DMSO or 5 nM actinomycin D for 4 h. Cells were immunostained with anti-HA antibody followed by Alexa Fluor 488 anti-mouse IgG and Flag antibodies followed by Alexa Fluor 546 anti-rabbit IgG. Representative immunofluorescence images were captured by confocal microscopy. ( d ) HEK293T cells were overexpressed with HA-RPS2 and His-ubiquitin and then treated with 10 μM MG132 for 4 h and 5 nM actinomycin D for indicated times. Ubiquitin conjugates were purified on Ni-NTA-agarose under denaturing conditions, and ubiquitinated RPS2 was detected by anti-HA antibodies. ( e ) U2OS cells were co-transfected with His-ubiquitin, HA-RPS2, and either siControl or siUSP47 and then treated with 10 μM MG132 and either DMSO or 5 nM actinomycin D for 4 h. Ubiquitin conjugates were purified on Ni-NTA agarose under denaturing conditions, and ubiquitinated RPS2 was detected by anti-HA antibodies. ( f ) U2OS cells transfected with siRNA against control or USP47 were treated with either DMSO or 5 nM actinomycin D for 4 h. Cell lysates were immunoblotted with the indicated antibodies. The uncropped blots and molecular weight markers are shown in Figure S7 .

    Journal: Cancers

    Article Title: USP47 Promotes Tumorigenesis by Negative Regulation of p53 through Deubiquitinating Ribosomal Protein S2

    doi: 10.3390/cancers12051137

    Figure Lengend Snippet: Ribosomal stress leads to RPS2 ubiquitination for MDM2 regulation to induce p53 by dissociation of USP47 and RPS2. ( a ) U2OS cells co-overexpressing either Flag vector or Flag-USP47 were treated with DMSO or 5 nM actinomycin D for 4 h, and immunoprecipitation was performed with anti-Flag agarose. Bound proteins were immunoblotted with the indicated antibodies. α-tubulin was used as a loading control. ( b ) HEK293T cells were co-overexpressed with HA-RPS2, Flag-USP47, and then treated with DMSO or 5 nM actinomycin D for 4 h. Then, cell lysates were immunoprecipitated with anti-Flag M2 agarose and detected with the indicated antibodies. ( c ) U2OS cells were co-overexpressed with Flag-USP47 and HA-RPS2 and then treated with DMSO or 5 nM actinomycin D for 4 h. Cells were immunostained with anti-HA antibody followed by Alexa Fluor 488 anti-mouse IgG and Flag antibodies followed by Alexa Fluor 546 anti-rabbit IgG. Representative immunofluorescence images were captured by confocal microscopy. ( d ) HEK293T cells were overexpressed with HA-RPS2 and His-ubiquitin and then treated with 10 μM MG132 for 4 h and 5 nM actinomycin D for indicated times. Ubiquitin conjugates were purified on Ni-NTA-agarose under denaturing conditions, and ubiquitinated RPS2 was detected by anti-HA antibodies. ( e ) U2OS cells were co-transfected with His-ubiquitin, HA-RPS2, and either siControl or siUSP47 and then treated with 10 μM MG132 and either DMSO or 5 nM actinomycin D for 4 h. Ubiquitin conjugates were purified on Ni-NTA agarose under denaturing conditions, and ubiquitinated RPS2 was detected by anti-HA antibodies. ( f ) U2OS cells transfected with siRNA against control or USP47 were treated with either DMSO or 5 nM actinomycin D for 4 h. Cell lysates were immunoblotted with the indicated antibodies. The uncropped blots and molecular weight markers are shown in Figure S7 .

    Article Snippet: Antibodies for HA, Myc, p53, and p21 were from Santa Cruz Biotechnology.

    Techniques: Plasmid Preparation, Immunoprecipitation, Immunofluorescence, Confocal Microscopy, Purification, Transfection, Molecular Weight

    USP47 regulates the ubiquitination of p53 and MDM2. ( a ) HEK293T cells were co-overexpressed with His-ubiquitin, p53, MDM2, HA-RPS2, myc-USP47, or myc-USP47 C109S for 24 h and then treated with MG132 (10 μM) for 4 h. Ubiquitin conjugates were purified on NiNTA-agarose under denaturing conditions, and ubiquitinated p53 was detected with anti-p53 antibody. ( b ) U2OS cells co-overexpressing His-ubiquitin, HA-RPS2, MDM2, Flag-USP47, or Flag-USP47 C109S as indicated were treated with MG132 (10 μM) for 4 h. Ubiquitin conjugates were purified on Ni-NTA agarose under denaturing conditions, and ubiquitinated MDM2 was detected with the anti-MDM2 antibody. The non-specific band is denoted by an asterisk. ( c ) H1299 cells were co-overexpressed with MDM2, His-ubiquitin, HA-RPS2, Flag-USP47, or Flag-USP47 C109S and then treated with 10 μM MG132 for 4 h. Ubiquitin conjugates were purified on Ni-NTA agarose under denaturing conditions, and ubiquitinated MDM2 was detected by anti-MDM2 antibodies. ( d ) HEK293T cells were overexpressed with MDM2, His-ubiquitin, and Flag-USP7 or Flag-USP47 and then treated with 10 μM MG132 for 4 h. Ubiquitin conjugates were purified on Ni-NTA agarose under denaturing conditions, and ubiquitinated MDM2 was detected by anti-MDM2 antibodies. The uncropped blots and molecular weight markers are shown in Figure S7 .

    Journal: Cancers

    Article Title: USP47 Promotes Tumorigenesis by Negative Regulation of p53 through Deubiquitinating Ribosomal Protein S2

    doi: 10.3390/cancers12051137

    Figure Lengend Snippet: USP47 regulates the ubiquitination of p53 and MDM2. ( a ) HEK293T cells were co-overexpressed with His-ubiquitin, p53, MDM2, HA-RPS2, myc-USP47, or myc-USP47 C109S for 24 h and then treated with MG132 (10 μM) for 4 h. Ubiquitin conjugates were purified on NiNTA-agarose under denaturing conditions, and ubiquitinated p53 was detected with anti-p53 antibody. ( b ) U2OS cells co-overexpressing His-ubiquitin, HA-RPS2, MDM2, Flag-USP47, or Flag-USP47 C109S as indicated were treated with MG132 (10 μM) for 4 h. Ubiquitin conjugates were purified on Ni-NTA agarose under denaturing conditions, and ubiquitinated MDM2 was detected with the anti-MDM2 antibody. The non-specific band is denoted by an asterisk. ( c ) H1299 cells were co-overexpressed with MDM2, His-ubiquitin, HA-RPS2, Flag-USP47, or Flag-USP47 C109S and then treated with 10 μM MG132 for 4 h. Ubiquitin conjugates were purified on Ni-NTA agarose under denaturing conditions, and ubiquitinated MDM2 was detected by anti-MDM2 antibodies. ( d ) HEK293T cells were overexpressed with MDM2, His-ubiquitin, and Flag-USP7 or Flag-USP47 and then treated with 10 μM MG132 for 4 h. Ubiquitin conjugates were purified on Ni-NTA agarose under denaturing conditions, and ubiquitinated MDM2 was detected by anti-MDM2 antibodies. The uncropped blots and molecular weight markers are shown in Figure S7 .

    Article Snippet: Antibodies for HA, Myc, p53, and p21 were from Santa Cruz Biotechnology.

    Techniques: Purification, Molecular Weight

    A schematic overview. Under normal conditions, USP47 deubiquitinates RPS2, and thus MDM2 inhibits p53 to maintain p53 protein levels. Under ribosomal stress, USP47 dissociates from RPS2, and thus ubiquitination of RPS2 is accumulated thereby inhibiting MDM2 to induce p53 protein levels for turning on the stress response signal.

    Journal: Cancers

    Article Title: USP47 Promotes Tumorigenesis by Negative Regulation of p53 through Deubiquitinating Ribosomal Protein S2

    doi: 10.3390/cancers12051137

    Figure Lengend Snippet: A schematic overview. Under normal conditions, USP47 deubiquitinates RPS2, and thus MDM2 inhibits p53 to maintain p53 protein levels. Under ribosomal stress, USP47 dissociates from RPS2, and thus ubiquitination of RPS2 is accumulated thereby inhibiting MDM2 to induce p53 protein levels for turning on the stress response signal.

    Article Snippet: Antibodies for HA, Myc, p53, and p21 were from Santa Cruz Biotechnology.

    Techniques:

    USP47 regulates p53 in an MDM2-dependent manner. ( a ) HeLa cells transfected with siRNA against control or USP47 were treated with either DMSO (control) or MG132 (10 μM) for 4 h. Cell lysates were immunoblotted with the indicated antibodies. β-actin was used as a loading control. The band intensity was measured by Image J and then normalized by β-actin. ( b ) U2OS cells overexpressed with Flag vector or Flag-USP47 were treated with either DMSO (control) or MG132 (10 μM) for 4 h. Cell lysates were immunoblotted with the indicated antibodies. ( c ) U2OS cells were overexpressed with increasing amounts of Myc-USP47 (0, 1, and 2 μg). After 24 h, cell lysates were immunoblotted with the indicated antibodies. ( d ) U2OS cells transfected with siRNA against control or USP47 were treated with 100 μg/mL cycloheximide and harvested at the indicated times. Cell lysates were immunoblotted with the indicated antibodies. HSP90 was used as a loading control. The band intensity was normalized by HSP90. ( e ) MDM2 +/+ p53 −/− and MDM2 −/− p53 −/− Mouse Embryonic Fibroblast (MEF) cell lines were co-overexpressed with p53, Myc-USP47, and then cell lysates were immunoblotted with the indicated antibodies. The non-specific band is denoted by an asterisk. ( f ) MDM2 +/+ p53 −/− and MDM2 −/− p53 −/− Mouse Embryonic Fibroblast (MEF) cell lines were overexpressed with p53 for 24 h and then transfected with siRNA against control or USP47 for 48 h. Cell lysates were immunoblotted with the indicated antibodies. The non-specific band is denoted by an asterisk. ( g ) The real-time PCR analysis of p53 downstream genes in A549 cells transfected with siRNA against control or USP47 for 48 h. Relative expression values are normalized against β-actin RNA levels, and graphs are shown as fold induction over control siRNA transfected cells. Data from three independent experiments represented by mean ± SD (* p

    Journal: Cancers

    Article Title: USP47 Promotes Tumorigenesis by Negative Regulation of p53 through Deubiquitinating Ribosomal Protein S2

    doi: 10.3390/cancers12051137

    Figure Lengend Snippet: USP47 regulates p53 in an MDM2-dependent manner. ( a ) HeLa cells transfected with siRNA against control or USP47 were treated with either DMSO (control) or MG132 (10 μM) for 4 h. Cell lysates were immunoblotted with the indicated antibodies. β-actin was used as a loading control. The band intensity was measured by Image J and then normalized by β-actin. ( b ) U2OS cells overexpressed with Flag vector or Flag-USP47 were treated with either DMSO (control) or MG132 (10 μM) for 4 h. Cell lysates were immunoblotted with the indicated antibodies. ( c ) U2OS cells were overexpressed with increasing amounts of Myc-USP47 (0, 1, and 2 μg). After 24 h, cell lysates were immunoblotted with the indicated antibodies. ( d ) U2OS cells transfected with siRNA against control or USP47 were treated with 100 μg/mL cycloheximide and harvested at the indicated times. Cell lysates were immunoblotted with the indicated antibodies. HSP90 was used as a loading control. The band intensity was normalized by HSP90. ( e ) MDM2 +/+ p53 −/− and MDM2 −/− p53 −/− Mouse Embryonic Fibroblast (MEF) cell lines were co-overexpressed with p53, Myc-USP47, and then cell lysates were immunoblotted with the indicated antibodies. The non-specific band is denoted by an asterisk. ( f ) MDM2 +/+ p53 −/− and MDM2 −/− p53 −/− Mouse Embryonic Fibroblast (MEF) cell lines were overexpressed with p53 for 24 h and then transfected with siRNA against control or USP47 for 48 h. Cell lysates were immunoblotted with the indicated antibodies. The non-specific band is denoted by an asterisk. ( g ) The real-time PCR analysis of p53 downstream genes in A549 cells transfected with siRNA against control or USP47 for 48 h. Relative expression values are normalized against β-actin RNA levels, and graphs are shown as fold induction over control siRNA transfected cells. Data from three independent experiments represented by mean ± SD (* p

    Article Snippet: Antibodies for HA, Myc, p53, and p21 were from Santa Cruz Biotechnology.

    Techniques: Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Expressing

    Suppression of USP47 inhibits cancer growth in vivo. ( a ) Western blot was conducted with tumor samples to evaluate the levels of USP47, p53, detected by indicated antibodies. ( b ) Immunofluorescence staining was conducted in dissected tumors to evaluate the levels of USP47, p53, MDM2 and detected by indicated antibodies. Immunofluorescence images were captured by confocal microscopy and quantified by ImageJ. Blue colors represent DAPI. Scale bars indicate 50 µm. ( c–d ) The tumor growth curves after initial treatment were shown; 2 mg/kg of siControl and siUSP47 were intratumorally administered into A549 xenografts every 3 days for 2 weeks. Tumor volumes were measured every 3 days after initial treatment. * p

    Journal: Cancers

    Article Title: USP47 Promotes Tumorigenesis by Negative Regulation of p53 through Deubiquitinating Ribosomal Protein S2

    doi: 10.3390/cancers12051137

    Figure Lengend Snippet: Suppression of USP47 inhibits cancer growth in vivo. ( a ) Western blot was conducted with tumor samples to evaluate the levels of USP47, p53, detected by indicated antibodies. ( b ) Immunofluorescence staining was conducted in dissected tumors to evaluate the levels of USP47, p53, MDM2 and detected by indicated antibodies. Immunofluorescence images were captured by confocal microscopy and quantified by ImageJ. Blue colors represent DAPI. Scale bars indicate 50 µm. ( c–d ) The tumor growth curves after initial treatment were shown; 2 mg/kg of siControl and siUSP47 were intratumorally administered into A549 xenografts every 3 days for 2 weeks. Tumor volumes were measured every 3 days after initial treatment. * p

    Article Snippet: Antibodies for HA, Myc, p53, and p21 were from Santa Cruz Biotechnology.

    Techniques: In Vivo, Western Blot, Immunofluorescence, Staining, Confocal Microscopy

    Inhibition of p53 transactivation function overcomes zinc deficiency-induced failure of neural tube closure. (A) Still frames from time-lapse live imaging of E8.5 Ven Myr embryos show that an inhibitor of p53 transactivation function, pifithrin-α, but not an inhibitor of p53 mitochondrial function, pifithrin-µ, can rescue TPEN-induced failure of neural tube closure. (B) Time to neural tube closure after TPEN+pifithrin-α ( n =6 from three independent experiments) relative to control ( n =6 from four independent experiments) (the neural tube never closes with TPEN treatment) ( n =6 from two independent experiments) or TPEN +pifithrin-µ-treated embryos ( n =5 from two independent experiments). (C-F) Representative still images (C) and quantitative data (D-F) of time-lapse imaging show the ability of pifithrin-α to rescue TPEN-induced failure of neural tube closure. In A and C, the white lines, red lines, yellow lines and purple line show the edges of neural fold. In D, the pink arrowhead indicates failure of neural tube closure; the dark gray and green arrowheads indicate the timepoint of neural tube closure. * P

    Journal: Development (Cambridge, England)

    Article Title: Zinc deficiency causes neural tube defects through attenuation of p53 ubiquitylation

    doi: 10.1242/dev.169797

    Figure Lengend Snippet: Inhibition of p53 transactivation function overcomes zinc deficiency-induced failure of neural tube closure. (A) Still frames from time-lapse live imaging of E8.5 Ven Myr embryos show that an inhibitor of p53 transactivation function, pifithrin-α, but not an inhibitor of p53 mitochondrial function, pifithrin-µ, can rescue TPEN-induced failure of neural tube closure. (B) Time to neural tube closure after TPEN+pifithrin-α ( n =6 from three independent experiments) relative to control ( n =6 from four independent experiments) (the neural tube never closes with TPEN treatment) ( n =6 from two independent experiments) or TPEN +pifithrin-µ-treated embryos ( n =5 from two independent experiments). (C-F) Representative still images (C) and quantitative data (D-F) of time-lapse imaging show the ability of pifithrin-α to rescue TPEN-induced failure of neural tube closure. In A and C, the white lines, red lines, yellow lines and purple line show the edges of neural fold. In D, the pink arrowhead indicates failure of neural tube closure; the dark gray and green arrowheads indicate the timepoint of neural tube closure. * P

    Article Snippet: Plasmids containing the coding region of human p53 ( TP53 -Myc-DDK-tagged) (RC200003, Origene) or human CHIP protein ( STUB1 -Myc-DDK-tagged) (RC200310, Origene) were transfected into cells using Xfect Transfection Reagent (Clontech) according to the manufacturer's protocol (1.5†µg or 2.5†µg plasmid for transfection per well of a 12-well plate; 0.5 or 1†µg for 24-well plate).

    Techniques: Inhibition, Imaging

    Model for the proposed mechanism of zinc deficiency-induced failure of neural tube closure. In the presence of zinc, Mdm2 protein can bind and ubiquitylate p53, resulting in p53 degradation, maintenance of cell survival and proper neural tube closure. Zinc deficiency attenuates Mdm2 binding and activity toward p53, resulting in p53 stabilization and an increase in p53 transcriptional activity, causing cell death and failure of neural tube closure.

    Journal: Development (Cambridge, England)

    Article Title: Zinc deficiency causes neural tube defects through attenuation of p53 ubiquitylation

    doi: 10.1242/dev.169797

    Figure Lengend Snippet: Model for the proposed mechanism of zinc deficiency-induced failure of neural tube closure. In the presence of zinc, Mdm2 protein can bind and ubiquitylate p53, resulting in p53 degradation, maintenance of cell survival and proper neural tube closure. Zinc deficiency attenuates Mdm2 binding and activity toward p53, resulting in p53 stabilization and an increase in p53 transcriptional activity, causing cell death and failure of neural tube closure.

    Article Snippet: Plasmids containing the coding region of human p53 ( TP53 -Myc-DDK-tagged) (RC200003, Origene) or human CHIP protein ( STUB1 -Myc-DDK-tagged) (RC200310, Origene) were transfected into cells using Xfect Transfection Reagent (Clontech) according to the manufacturer's protocol (1.5†µg or 2.5†µg plasmid for transfection per well of a 12-well plate; 0.5 or 1†µg for 24-well plate).

    Techniques: Binding Assay, Activity Assay

    Zinc deficiency attenuates p53 ubiquitylation. (A) Schematic of the regulation of p53 ubiquitylation by the zinc-regulated E3 ubiquitin ligase Mdm2 and by the p53-deubiquitylating enzyme Hausp, leading to p53 degradation or stabilization. The N terminus of Mdm2 binds p53 via a TAZ domain, whereas the C-terminal zinc-regulated RING domain is required for ubiquitin ligase activity towards p53. TAZ, transactive zone. (B) E9.5 primary neuroepithelial cells treated with MG132 proteasome inhibitor and TPEN or TPEN+ZnSO 4 , followed by p53 co-immunoprecipitation and western blot analysis with antibodies against ubiquitin and Hausp shows TPEN decreases p53 ubiquitylation and Hausp binding. Graph shows western blot quantification data. Immunoprecipitated p53 is used as a loading control. Data are mean±s.d. All treatments were normalized to control. * P

    Journal: Development (Cambridge, England)

    Article Title: Zinc deficiency causes neural tube defects through attenuation of p53 ubiquitylation

    doi: 10.1242/dev.169797

    Figure Lengend Snippet: Zinc deficiency attenuates p53 ubiquitylation. (A) Schematic of the regulation of p53 ubiquitylation by the zinc-regulated E3 ubiquitin ligase Mdm2 and by the p53-deubiquitylating enzyme Hausp, leading to p53 degradation or stabilization. The N terminus of Mdm2 binds p53 via a TAZ domain, whereas the C-terminal zinc-regulated RING domain is required for ubiquitin ligase activity towards p53. TAZ, transactive zone. (B) E9.5 primary neuroepithelial cells treated with MG132 proteasome inhibitor and TPEN or TPEN+ZnSO 4 , followed by p53 co-immunoprecipitation and western blot analysis with antibodies against ubiquitin and Hausp shows TPEN decreases p53 ubiquitylation and Hausp binding. Graph shows western blot quantification data. Immunoprecipitated p53 is used as a loading control. Data are mean±s.d. All treatments were normalized to control. * P

    Article Snippet: Plasmids containing the coding region of human p53 ( TP53 -Myc-DDK-tagged) (RC200003, Origene) or human CHIP protein ( STUB1 -Myc-DDK-tagged) (RC200310, Origene) were transfected into cells using Xfect Transfection Reagent (Clontech) according to the manufacturer's protocol (1.5†µg or 2.5†µg plasmid for transfection per well of a 12-well plate; 0.5 or 1†µg for 24-well plate).

    Techniques: Activity Assay, Immunoprecipitation, Western Blot, Binding Assay

    p53 plays a crucial role in zinc deficiency-induced apoptosis. Western blot analysis of brain tissue from E9.5 embryos (each sample was pooled from four or five embryos, three independent experiments were performed) (A) and E9.5 primary neuroepithelial cells (from three independent experiments) (B) demonstrate that p53 levels increased in both the cytoplasm and nucleus after TPEN treatment for the indicated length of time, and p53 levels in both cellular compartments were reduced by concurrent zinc supplementation. Graphs show the corresponding quantification results of western blots. Data are mean±s.d. * P

    Journal: Development (Cambridge, England)

    Article Title: Zinc deficiency causes neural tube defects through attenuation of p53 ubiquitylation

    doi: 10.1242/dev.169797

    Figure Lengend Snippet: p53 plays a crucial role in zinc deficiency-induced apoptosis. Western blot analysis of brain tissue from E9.5 embryos (each sample was pooled from four or five embryos, three independent experiments were performed) (A) and E9.5 primary neuroepithelial cells (from three independent experiments) (B) demonstrate that p53 levels increased in both the cytoplasm and nucleus after TPEN treatment for the indicated length of time, and p53 levels in both cellular compartments were reduced by concurrent zinc supplementation. Graphs show the corresponding quantification results of western blots. Data are mean±s.d. * P

    Article Snippet: Plasmids containing the coding region of human p53 ( TP53 -Myc-DDK-tagged) (RC200003, Origene) or human CHIP protein ( STUB1 -Myc-DDK-tagged) (RC200310, Origene) were transfected into cells using Xfect Transfection Reagent (Clontech) according to the manufacturer's protocol (1.5†µg or 2.5†µg plasmid for transfection per well of a 12-well plate; 0.5 or 1†µg for 24-well plate).

    Techniques: Western Blot

    Representative images of Immunohistochemistry for molecular markers of glioblastoma. Images for Ki67 (high above and low below), p53 (positive staining above and negative below), and IDH1 (positive staining above and negative below) are presented.

    Journal: Scientific Reports

    Article Title: Methylation of MGMT promoter does not predict response to temozolomide in patients with glioblastoma in Donostia Hospital

    doi: 10.1038/s41598-020-75477-9

    Figure Lengend Snippet: Representative images of Immunohistochemistry for molecular markers of glioblastoma. Images for Ki67 (high above and low below), p53 (positive staining above and negative below), and IDH1 (positive staining above and negative below) are presented.

    Article Snippet: The following primary antibodies were used: Ki67 (Roche; Ref 790-4286, dilution 1:100), p53 (Roche; Ref 790-2912, predilution 1:20), IDH1 (vitro master diagnostica; Ref R132H antibody H09, predilution) and ATRX (Sigma-Aldrich; Ref HPA001906, 1:200).

    Techniques: Immunohistochemistry, Staining