ck1δ antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ck1δ antibody
    CK1ε and <t>CK1δ</t> are upregulated in colon cancer and associated with poor clinical outcome. (A, B) CK1ε and CK1δ mRNA were detected in colon cancer and paired adjacent normal tissues by using qRT-PCR. (C, D) CK1ε and CK1δ proteins were detected in colon cancer and paired adjacent normal tissues by using IHC. (E) Kaplan-Meier analysis of the CK1ε and CK1δ expression on overall survival of colon cancer patients (** p <0.01).
    Ck1δ Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
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    ck1δ antibody - by Bioz Stars, 2023-02
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    1) Product Images from "IC261, a specific inhibitor of CK1δ/ε, promotes aerobic glycolysis through p53-dependent mechanisms in colon cancer"

    Article Title: IC261, a specific inhibitor of CK1δ/ε, promotes aerobic glycolysis through p53-dependent mechanisms in colon cancer

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.40960

    CK1ε and CK1δ are upregulated in colon cancer and associated with poor clinical outcome. (A, B) CK1ε and CK1δ mRNA were detected in colon cancer and paired adjacent normal tissues by using qRT-PCR. (C, D) CK1ε and CK1δ proteins were detected in colon cancer and paired adjacent normal tissues by using IHC. (E) Kaplan-Meier analysis of the CK1ε and CK1δ expression on overall survival of colon cancer patients (** p <0.01).
    Figure Legend Snippet: CK1ε and CK1δ are upregulated in colon cancer and associated with poor clinical outcome. (A, B) CK1ε and CK1δ mRNA were detected in colon cancer and paired adjacent normal tissues by using qRT-PCR. (C, D) CK1ε and CK1δ proteins were detected in colon cancer and paired adjacent normal tissues by using IHC. (E) Kaplan-Meier analysis of the CK1ε and CK1δ expression on overall survival of colon cancer patients (** p <0.01).

    Techniques Used: Quantitative RT-PCR, Expressing

    Correlation between the clinicopathological features and  CK1δ  expression
    Figure Legend Snippet: Correlation between the clinicopathological features and CK1δ expression

    Techniques Used: Expressing

    ck1δ antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ck1δ antibody
    CK1ε and <t>CK1δ</t> are upregulated in colon cancer and associated with poor clinical outcome. (A, B) CK1ε and CK1δ mRNA were detected in colon cancer and paired adjacent normal tissues by using qRT-PCR. (C, D) CK1ε and CK1δ proteins were detected in colon cancer and paired adjacent normal tissues by using IHC. (E) Kaplan-Meier analysis of the CK1ε and CK1δ expression on overall survival of colon cancer patients (** p <0.01).
    Ck1δ Antibody, supplied by Cell Signaling Technology Inc, 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/ck1δ antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ck1δ antibody - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "IC261, a specific inhibitor of CK1δ/ε, promotes aerobic glycolysis through p53-dependent mechanisms in colon cancer"

    Article Title: IC261, a specific inhibitor of CK1δ/ε, promotes aerobic glycolysis through p53-dependent mechanisms in colon cancer

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.40960

    CK1ε and CK1δ are upregulated in colon cancer and associated with poor clinical outcome. (A, B) CK1ε and CK1δ mRNA were detected in colon cancer and paired adjacent normal tissues by using qRT-PCR. (C, D) CK1ε and CK1δ proteins were detected in colon cancer and paired adjacent normal tissues by using IHC. (E) Kaplan-Meier analysis of the CK1ε and CK1δ expression on overall survival of colon cancer patients (** p <0.01).
    Figure Legend Snippet: CK1ε and CK1δ are upregulated in colon cancer and associated with poor clinical outcome. (A, B) CK1ε and CK1δ mRNA were detected in colon cancer and paired adjacent normal tissues by using qRT-PCR. (C, D) CK1ε and CK1δ proteins were detected in colon cancer and paired adjacent normal tissues by using IHC. (E) Kaplan-Meier analysis of the CK1ε and CK1δ expression on overall survival of colon cancer patients (** p <0.01).

    Techniques Used: Quantitative RT-PCR, Expressing

    Correlation between the clinicopathological features and  CK1δ  expression
    Figure Legend Snippet: Correlation between the clinicopathological features and CK1δ expression

    Techniques Used: Expressing

    ck1δ  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
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    Cell Signaling Technology Inc ck1δ
    a HEK293 cells were transfected with indicated plasmids, followed by immunoprecipitation with FLAG-agarose beads and immunoblotting. b HEK293 cells were transfected with various FLAG-CK1 isoforms, immunoprecipitated with FLAG-agarose beads and analyzed by immunoblotting. c Endogenous <t>CK1δ</t> protein in SUM159 cell lysates were pulled down with anti-S CK1δ antibody, and associated ASCT2 were detected by immunoblotting with anti-ASCT2 Ab (top, IP). Cell lysates were subjected to immunoblotting (bottom, input). d MDA-MB-231 cells were transfected with indicated CK1 isoforms, then analyzed by immunoblotting. e MDA-MB-231 cells were transfected siRNA targeting CK1δ for 48 h, followed by immunoblotting. f HEK293 cells were transfected with FLAG-SPOP and HA-ASCT2 and treated with D4476 for 12 h before harvesting, followed by immunoprecipitation with FLAG-agarose beads and analyzed by immunoblotting. g MDA-MB-231 cells were treated with different concentrations of CK1 inhibitor D4476 for 24 h, then analyzed by immunoblotting. h , i MDA-MB-231 cells were transfected with si-NC or siRNA targeting CK1δ (si-CK1δ#1), then incubated with CHX for various time points, and analyzed by immunoblotting ies ( h ), the band density of ASCT2 was quantified using ImageJ software and normalized to α-tubulin (mean ± SD, n = 3) ( i ). j HEK293 cells were transfected with indicated plasmids, lysed under denaturing conditions, followed by Ni-beads pulldown, and immunoblotting for ASCT2. Source data are provided as a Source Data file.
    Ck1δ, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ck1δ - by Bioz Stars, 2023-02
    94/100 stars

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    1) Product Images from "Neddylation inhibition induces glutamine uptake and metabolism by targeting CRL3 SPOP E3 ligase in cancer cells"

    Article Title: Neddylation inhibition induces glutamine uptake and metabolism by targeting CRL3 SPOP E3 ligase in cancer cells

    Journal: Nature Communications

    doi: 10.1038/s41467-022-30559-2

    a HEK293 cells were transfected with indicated plasmids, followed by immunoprecipitation with FLAG-agarose beads and immunoblotting. b HEK293 cells were transfected with various FLAG-CK1 isoforms, immunoprecipitated with FLAG-agarose beads and analyzed by immunoblotting. c Endogenous CK1δ protein in SUM159 cell lysates were pulled down with anti-S CK1δ antibody, and associated ASCT2 were detected by immunoblotting with anti-ASCT2 Ab (top, IP). Cell lysates were subjected to immunoblotting (bottom, input). d MDA-MB-231 cells were transfected with indicated CK1 isoforms, then analyzed by immunoblotting. e MDA-MB-231 cells were transfected siRNA targeting CK1δ for 48 h, followed by immunoblotting. f HEK293 cells were transfected with FLAG-SPOP and HA-ASCT2 and treated with D4476 for 12 h before harvesting, followed by immunoprecipitation with FLAG-agarose beads and analyzed by immunoblotting. g MDA-MB-231 cells were treated with different concentrations of CK1 inhibitor D4476 for 24 h, then analyzed by immunoblotting. h , i MDA-MB-231 cells were transfected with si-NC or siRNA targeting CK1δ (si-CK1δ#1), then incubated with CHX for various time points, and analyzed by immunoblotting ies ( h ), the band density of ASCT2 was quantified using ImageJ software and normalized to α-tubulin (mean ± SD, n = 3) ( i ). j HEK293 cells were transfected with indicated plasmids, lysed under denaturing conditions, followed by Ni-beads pulldown, and immunoblotting for ASCT2. Source data are provided as a Source Data file.
    Figure Legend Snippet: a HEK293 cells were transfected with indicated plasmids, followed by immunoprecipitation with FLAG-agarose beads and immunoblotting. b HEK293 cells were transfected with various FLAG-CK1 isoforms, immunoprecipitated with FLAG-agarose beads and analyzed by immunoblotting. c Endogenous CK1δ protein in SUM159 cell lysates were pulled down with anti-S CK1δ antibody, and associated ASCT2 were detected by immunoblotting with anti-ASCT2 Ab (top, IP). Cell lysates were subjected to immunoblotting (bottom, input). d MDA-MB-231 cells were transfected with indicated CK1 isoforms, then analyzed by immunoblotting. e MDA-MB-231 cells were transfected siRNA targeting CK1δ for 48 h, followed by immunoblotting. f HEK293 cells were transfected with FLAG-SPOP and HA-ASCT2 and treated with D4476 for 12 h before harvesting, followed by immunoprecipitation with FLAG-agarose beads and analyzed by immunoblotting. g MDA-MB-231 cells were treated with different concentrations of CK1 inhibitor D4476 for 24 h, then analyzed by immunoblotting. h , i MDA-MB-231 cells were transfected with si-NC or siRNA targeting CK1δ (si-CK1δ#1), then incubated with CHX for various time points, and analyzed by immunoblotting ies ( h ), the band density of ASCT2 was quantified using ImageJ software and normalized to α-tubulin (mean ± SD, n = 3) ( i ). j HEK293 cells were transfected with indicated plasmids, lysed under denaturing conditions, followed by Ni-beads pulldown, and immunoblotting for ASCT2. Source data are provided as a Source Data file.

    Techniques Used: Transfection, Immunoprecipitation, Western Blot, Incubation, Software

    anti lats1 3477  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti lats1 3477
    Cullin3 SPOP is the physiological E3 ubiquitin ligase for <t>LATS1.</t> (a) IB analysis of WCLs derived from 293T, 786-O and A498 cells treated with 10μM MG132 for 10 hr. (b) IB analysis of immunoprecipitates (IPs) and WCLs derived from 293T kidney cells transfected with indicated constructs. Cells were treated with MG132 (10 μM) before harvesting. (c) IB analysis of WCLs derived from A498 cells transfected with Cullin3 siRNA. (d) IB analysis of WCLs derived from 786-O cells after the specified duration of 100 μg/ml cycloheximide (CHX) transfected with Cullin3 siRNA. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated constructs. Cells were treated with MG132 (10μM) before harvesting. (g) IB analysis of WCLs derived from 293T kidney cells transfected with increasing doses of plasmid encoding SPOP. (h) IB analysis of WCLs derived from 786-O cells transfected with increasing doses of plasmid encoding SPOP. (i) IB analysis of WCLs derived from 293T kidney cells or A498 cells transfected with indicated constructs. Where indicated, cells were treated with 10μM MG132 before harvesting. (j) IB analysis of WCLs and products of ubiquitination derived from 293T cells transfected with indicated constructs.
    Anti Lats1 3477, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti lats1 3477 - by Bioz Stars, 2023-02
    94/100 stars

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    1) Product Images from "SPOP promotes ubiquitination and degradation of LATS1 to enhance kidney cancer progression"

    Article Title: SPOP promotes ubiquitination and degradation of LATS1 to enhance kidney cancer progression

    Journal: EBioMedicine

    doi: 10.1016/j.ebiom.2020.102795

    Cullin3 SPOP is the physiological E3 ubiquitin ligase for LATS1. (a) IB analysis of WCLs derived from 293T, 786-O and A498 cells treated with 10μM MG132 for 10 hr. (b) IB analysis of immunoprecipitates (IPs) and WCLs derived from 293T kidney cells transfected with indicated constructs. Cells were treated with MG132 (10 μM) before harvesting. (c) IB analysis of WCLs derived from A498 cells transfected with Cullin3 siRNA. (d) IB analysis of WCLs derived from 786-O cells after the specified duration of 100 μg/ml cycloheximide (CHX) transfected with Cullin3 siRNA. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated constructs. Cells were treated with MG132 (10μM) before harvesting. (g) IB analysis of WCLs derived from 293T kidney cells transfected with increasing doses of plasmid encoding SPOP. (h) IB analysis of WCLs derived from 786-O cells transfected with increasing doses of plasmid encoding SPOP. (i) IB analysis of WCLs derived from 293T kidney cells or A498 cells transfected with indicated constructs. Where indicated, cells were treated with 10μM MG132 before harvesting. (j) IB analysis of WCLs and products of ubiquitination derived from 293T cells transfected with indicated constructs.
    Figure Legend Snippet: Cullin3 SPOP is the physiological E3 ubiquitin ligase for LATS1. (a) IB analysis of WCLs derived from 293T, 786-O and A498 cells treated with 10μM MG132 for 10 hr. (b) IB analysis of immunoprecipitates (IPs) and WCLs derived from 293T kidney cells transfected with indicated constructs. Cells were treated with MG132 (10 μM) before harvesting. (c) IB analysis of WCLs derived from A498 cells transfected with Cullin3 siRNA. (d) IB analysis of WCLs derived from 786-O cells after the specified duration of 100 μg/ml cycloheximide (CHX) transfected with Cullin3 siRNA. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated constructs. Cells were treated with MG132 (10μM) before harvesting. (g) IB analysis of WCLs derived from 293T kidney cells transfected with increasing doses of plasmid encoding SPOP. (h) IB analysis of WCLs derived from 786-O cells transfected with increasing doses of plasmid encoding SPOP. (i) IB analysis of WCLs derived from 293T kidney cells or A498 cells transfected with indicated constructs. Where indicated, cells were treated with 10μM MG132 before harvesting. (j) IB analysis of WCLs and products of ubiquitination derived from 293T cells transfected with indicated constructs.

    Techniques Used: Derivative Assay, Transfection, Construct, Plasmid Preparation

    Cullin3 SPOP E3 ubiquitin ligase negatively regulates the protein stability of LATS1. (a) IB analysis of WCLs derived from A498 or 786-O cells infected with the indicated lentiviral shRNA vectors. (b) IB analysis of WCLs derived from 293T or A498 cells transfected with HA-tagged SPOP plasmid. (c) Quantitative real-time PCR (qRT-PCR) analysis to detect LATS1 and SPOP mRNA levels derived from 786-O or A498 cells after depletion of SPOP. Data are shown as mean ±SD of three independent experiments. * P <0.05, Student's t test. (d) IB analysis of WCLs derived from 293T cells after the specified duration of cycloheximide (CHX) transfected with Myc-tagged SPOP plasmid. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs derived from A498 cells after the specified duration of 100μg/ml cycloheximide (CHX) infected with indicated lentiviral shRNA vectors. (g) The abundance of LATS1 protein in (f) was quantified and plotted.
    Figure Legend Snippet: Cullin3 SPOP E3 ubiquitin ligase negatively regulates the protein stability of LATS1. (a) IB analysis of WCLs derived from A498 or 786-O cells infected with the indicated lentiviral shRNA vectors. (b) IB analysis of WCLs derived from 293T or A498 cells transfected with HA-tagged SPOP plasmid. (c) Quantitative real-time PCR (qRT-PCR) analysis to detect LATS1 and SPOP mRNA levels derived from 786-O or A498 cells after depletion of SPOP. Data are shown as mean ±SD of three independent experiments. * P <0.05, Student's t test. (d) IB analysis of WCLs derived from 293T cells after the specified duration of cycloheximide (CHX) transfected with Myc-tagged SPOP plasmid. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs derived from A498 cells after the specified duration of 100μg/ml cycloheximide (CHX) infected with indicated lentiviral shRNA vectors. (g) The abundance of LATS1 protein in (f) was quantified and plotted.

    Techniques Used: Derivative Assay, Infection, shRNA, Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    SPOP-mediated ubiquitination and degradation of LATS1 depends on the degron motif. (a) Amino acid sequence alignment of LATS1 with the SPOP–binding motif (degron) in known substrates of SPOP. (b) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (c) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting. (d) IB analysis of WCLs derived from 293T cells after the specified duration of 100μg/ml cycloheximide (CHX) transfected with indicated Flag-tagged LATS1 plasmids. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 hr before harvesting. (g) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting. (h) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (i) IB analysis of WCLs derived from 293T cells after the specified duration of 100μg/ml cycloheximide (CHX) transfected with indicated Flag-tagged LATS1 plasmids. (j) The abundance of LATS1 protein in (i) was quantified and plotted.
    Figure Legend Snippet: SPOP-mediated ubiquitination and degradation of LATS1 depends on the degron motif. (a) Amino acid sequence alignment of LATS1 with the SPOP–binding motif (degron) in known substrates of SPOP. (b) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (c) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting. (d) IB analysis of WCLs derived from 293T cells after the specified duration of 100μg/ml cycloheximide (CHX) transfected with indicated Flag-tagged LATS1 plasmids. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 hr before harvesting. (g) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting. (h) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (i) IB analysis of WCLs derived from 293T cells after the specified duration of 100μg/ml cycloheximide (CHX) transfected with indicated Flag-tagged LATS1 plasmids. (j) The abundance of LATS1 protein in (i) was quantified and plotted.

    Techniques Used: Sequencing, Binding Assay, Derivative Assay, Transfection

    CKΙδ promotes the interaction and degradation of LATS1 by SPOP. (a) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (b) IB analysis of WCLs derived from 293T, A498 and 786-O cells incubated with CKΙ inhibitor IC261 (50μM) or D4476 (20μM) before harvesting. (c) IB analysis of WCLs derived from kidney derived 293T cells transfected with CKΙδ siRNA. (d) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T kidney cells transfected with indicated plasmids and treated with 10μM MG132 for 10 hr before harvesting. (e) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (f) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (g) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting.
    Figure Legend Snippet: CKΙδ promotes the interaction and degradation of LATS1 by SPOP. (a) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (b) IB analysis of WCLs derived from 293T, A498 and 786-O cells incubated with CKΙ inhibitor IC261 (50μM) or D4476 (20μM) before harvesting. (c) IB analysis of WCLs derived from kidney derived 293T cells transfected with CKΙδ siRNA. (d) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T kidney cells transfected with indicated plasmids and treated with 10μM MG132 for 10 hr before harvesting. (e) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (f) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (g) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting.

    Techniques Used: Derivative Assay, Transfection, Incubation

    SPOP promotes tumorigenesis. (a) 786-O-pcDNA3.1, 786-O-SPOP polyclonal stable kidney cancer cell lines were injected subcutaneously into the BALB/c-nu/nu mice. 40 days later, mice were anesthetic and taken a picture. (b) The tumours were dissected and taken a picture. (c) The weights of the dissected tumours in (b). (d) In vivo tumour growth was measured over the indicated time period. (e) The body weights of the BALB/c-nu/nu mice are measured over the indicated time period. (f) Immunohistochemistry staining of SPOP and LATS1 in tissue sections of xenografted tumours in BALB/c-nu/nu mice injected with the indicated cell lines. Scale bars, 20 μm. (g) IB analysis of the LATS1 protein levels in the dissected tumours. (h) Statistical analyses of the correlation between SPOP and LATS1 expression in cytoplasm of the kidney cancer tissue microarray. (i) Representative images of SPOP and LATS1 IHC from 89 cases of kidney cancer.
    Figure Legend Snippet: SPOP promotes tumorigenesis. (a) 786-O-pcDNA3.1, 786-O-SPOP polyclonal stable kidney cancer cell lines were injected subcutaneously into the BALB/c-nu/nu mice. 40 days later, mice were anesthetic and taken a picture. (b) The tumours were dissected and taken a picture. (c) The weights of the dissected tumours in (b). (d) In vivo tumour growth was measured over the indicated time period. (e) The body weights of the BALB/c-nu/nu mice are measured over the indicated time period. (f) Immunohistochemistry staining of SPOP and LATS1 in tissue sections of xenografted tumours in BALB/c-nu/nu mice injected with the indicated cell lines. Scale bars, 20 μm. (g) IB analysis of the LATS1 protein levels in the dissected tumours. (h) Statistical analyses of the correlation between SPOP and LATS1 expression in cytoplasm of the kidney cancer tissue microarray. (i) Representative images of SPOP and LATS1 IHC from 89 cases of kidney cancer.

    Techniques Used: Injection, In Vivo, Immunohistochemistry, Staining, Expressing, Microarray

    anti ck1 12417 antibodies  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti ck1 12417 antibodies
    Anti Ck1 12417 Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ck1δ  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ck1δ
    Ck1δ, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti ck1δ  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti ck1δ
    Anti Ck1δ, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti ck1δ  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti ck1δ
    Anti Ck1δ, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti ck1δ  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti ck1δ
    Anti Ck1δ, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc ck1δ antibody
    CK1ε and <t>CK1δ</t> are upregulated in colon cancer and associated with poor clinical outcome. (A, B) CK1ε and CK1δ mRNA were detected in colon cancer and paired adjacent normal tissues by using qRT-PCR. (C, D) CK1ε and CK1δ proteins were detected in colon cancer and paired adjacent normal tissues by using IHC. (E) Kaplan-Meier analysis of the CK1ε and CK1δ expression on overall survival of colon cancer patients (** p <0.01).
    Ck1δ Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc ck1δ
    a HEK293 cells were transfected with indicated plasmids, followed by immunoprecipitation with FLAG-agarose beads and immunoblotting. b HEK293 cells were transfected with various FLAG-CK1 isoforms, immunoprecipitated with FLAG-agarose beads and analyzed by immunoblotting. c Endogenous <t>CK1δ</t> protein in SUM159 cell lysates were pulled down with anti-S CK1δ antibody, and associated ASCT2 were detected by immunoblotting with anti-ASCT2 Ab (top, IP). Cell lysates were subjected to immunoblotting (bottom, input). d MDA-MB-231 cells were transfected with indicated CK1 isoforms, then analyzed by immunoblotting. e MDA-MB-231 cells were transfected siRNA targeting CK1δ for 48 h, followed by immunoblotting. f HEK293 cells were transfected with FLAG-SPOP and HA-ASCT2 and treated with D4476 for 12 h before harvesting, followed by immunoprecipitation with FLAG-agarose beads and analyzed by immunoblotting. g MDA-MB-231 cells were treated with different concentrations of CK1 inhibitor D4476 for 24 h, then analyzed by immunoblotting. h , i MDA-MB-231 cells were transfected with si-NC or siRNA targeting CK1δ (si-CK1δ#1), then incubated with CHX for various time points, and analyzed by immunoblotting ies ( h ), the band density of ASCT2 was quantified using ImageJ software and normalized to α-tubulin (mean ± SD, n = 3) ( i ). j HEK293 cells were transfected with indicated plasmids, lysed under denaturing conditions, followed by Ni-beads pulldown, and immunoblotting for ASCT2. Source data are provided as a Source Data file.
    Ck1δ, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti lats1 3477
    Cullin3 SPOP is the physiological E3 ubiquitin ligase for <t>LATS1.</t> (a) IB analysis of WCLs derived from 293T, 786-O and A498 cells treated with 10μM MG132 for 10 hr. (b) IB analysis of immunoprecipitates (IPs) and WCLs derived from 293T kidney cells transfected with indicated constructs. Cells were treated with MG132 (10 μM) before harvesting. (c) IB analysis of WCLs derived from A498 cells transfected with Cullin3 siRNA. (d) IB analysis of WCLs derived from 786-O cells after the specified duration of 100 μg/ml cycloheximide (CHX) transfected with Cullin3 siRNA. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated constructs. Cells were treated with MG132 (10μM) before harvesting. (g) IB analysis of WCLs derived from 293T kidney cells transfected with increasing doses of plasmid encoding SPOP. (h) IB analysis of WCLs derived from 786-O cells transfected with increasing doses of plasmid encoding SPOP. (i) IB analysis of WCLs derived from 293T kidney cells or A498 cells transfected with indicated constructs. Where indicated, cells were treated with 10μM MG132 before harvesting. (j) IB analysis of WCLs and products of ubiquitination derived from 293T cells transfected with indicated constructs.
    Anti Lats1 3477, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti ck1 12417 antibodies
    Cullin3 SPOP is the physiological E3 ubiquitin ligase for <t>LATS1.</t> (a) IB analysis of WCLs derived from 293T, 786-O and A498 cells treated with 10μM MG132 for 10 hr. (b) IB analysis of immunoprecipitates (IPs) and WCLs derived from 293T kidney cells transfected with indicated constructs. Cells were treated with MG132 (10 μM) before harvesting. (c) IB analysis of WCLs derived from A498 cells transfected with Cullin3 siRNA. (d) IB analysis of WCLs derived from 786-O cells after the specified duration of 100 μg/ml cycloheximide (CHX) transfected with Cullin3 siRNA. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated constructs. Cells were treated with MG132 (10μM) before harvesting. (g) IB analysis of WCLs derived from 293T kidney cells transfected with increasing doses of plasmid encoding SPOP. (h) IB analysis of WCLs derived from 786-O cells transfected with increasing doses of plasmid encoding SPOP. (i) IB analysis of WCLs derived from 293T kidney cells or A498 cells transfected with indicated constructs. Where indicated, cells were treated with 10μM MG132 before harvesting. (j) IB analysis of WCLs and products of ubiquitination derived from 293T cells transfected with indicated constructs.
    Anti Ck1 12417 Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti ck1δ
    Cullin3 SPOP is the physiological E3 ubiquitin ligase for <t>LATS1.</t> (a) IB analysis of WCLs derived from 293T, 786-O and A498 cells treated with 10μM MG132 for 10 hr. (b) IB analysis of immunoprecipitates (IPs) and WCLs derived from 293T kidney cells transfected with indicated constructs. Cells were treated with MG132 (10 μM) before harvesting. (c) IB analysis of WCLs derived from A498 cells transfected with Cullin3 siRNA. (d) IB analysis of WCLs derived from 786-O cells after the specified duration of 100 μg/ml cycloheximide (CHX) transfected with Cullin3 siRNA. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated constructs. Cells were treated with MG132 (10μM) before harvesting. (g) IB analysis of WCLs derived from 293T kidney cells transfected with increasing doses of plasmid encoding SPOP. (h) IB analysis of WCLs derived from 786-O cells transfected with increasing doses of plasmid encoding SPOP. (i) IB analysis of WCLs derived from 293T kidney cells or A498 cells transfected with indicated constructs. Where indicated, cells were treated with 10μM MG132 before harvesting. (j) IB analysis of WCLs and products of ubiquitination derived from 293T cells transfected with indicated constructs.
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    Image Search Results


    CK1ε and CK1δ are upregulated in colon cancer and associated with poor clinical outcome. (A, B) CK1ε and CK1δ mRNA were detected in colon cancer and paired adjacent normal tissues by using qRT-PCR. (C, D) CK1ε and CK1δ proteins were detected in colon cancer and paired adjacent normal tissues by using IHC. (E) Kaplan-Meier analysis of the CK1ε and CK1δ expression on overall survival of colon cancer patients (** p <0.01).

    Journal: International Journal of Biological Sciences

    Article Title: IC261, a specific inhibitor of CK1δ/ε, promotes aerobic glycolysis through p53-dependent mechanisms in colon cancer

    doi: 10.7150/ijbs.40960

    Figure Lengend Snippet: CK1ε and CK1δ are upregulated in colon cancer and associated with poor clinical outcome. (A, B) CK1ε and CK1δ mRNA were detected in colon cancer and paired adjacent normal tissues by using qRT-PCR. (C, D) CK1ε and CK1δ proteins were detected in colon cancer and paired adjacent normal tissues by using IHC. (E) Kaplan-Meier analysis of the CK1ε and CK1δ expression on overall survival of colon cancer patients (** p <0.01).

    Article Snippet: The slides were incubated overnight at 4°C with CK1ε and CK1δ antibody (Cell Signaling Technology USA), followed by incubation with secondary antibodies (Abm, China).

    Techniques: Quantitative RT-PCR, Expressing

    Correlation between the clinicopathological features and  CK1δ  expression

    Journal: International Journal of Biological Sciences

    Article Title: IC261, a specific inhibitor of CK1δ/ε, promotes aerobic glycolysis through p53-dependent mechanisms in colon cancer

    doi: 10.7150/ijbs.40960

    Figure Lengend Snippet: Correlation between the clinicopathological features and CK1δ expression

    Article Snippet: The slides were incubated overnight at 4°C with CK1ε and CK1δ antibody (Cell Signaling Technology USA), followed by incubation with secondary antibodies (Abm, China).

    Techniques: Expressing

    a HEK293 cells were transfected with indicated plasmids, followed by immunoprecipitation with FLAG-agarose beads and immunoblotting. b HEK293 cells were transfected with various FLAG-CK1 isoforms, immunoprecipitated with FLAG-agarose beads and analyzed by immunoblotting. c Endogenous CK1δ protein in SUM159 cell lysates were pulled down with anti-S CK1δ antibody, and associated ASCT2 were detected by immunoblotting with anti-ASCT2 Ab (top, IP). Cell lysates were subjected to immunoblotting (bottom, input). d MDA-MB-231 cells were transfected with indicated CK1 isoforms, then analyzed by immunoblotting. e MDA-MB-231 cells were transfected siRNA targeting CK1δ for 48 h, followed by immunoblotting. f HEK293 cells were transfected with FLAG-SPOP and HA-ASCT2 and treated with D4476 for 12 h before harvesting, followed by immunoprecipitation with FLAG-agarose beads and analyzed by immunoblotting. g MDA-MB-231 cells were treated with different concentrations of CK1 inhibitor D4476 for 24 h, then analyzed by immunoblotting. h , i MDA-MB-231 cells were transfected with si-NC or siRNA targeting CK1δ (si-CK1δ#1), then incubated with CHX for various time points, and analyzed by immunoblotting ies ( h ), the band density of ASCT2 was quantified using ImageJ software and normalized to α-tubulin (mean ± SD, n = 3) ( i ). j HEK293 cells were transfected with indicated plasmids, lysed under denaturing conditions, followed by Ni-beads pulldown, and immunoblotting for ASCT2. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Neddylation inhibition induces glutamine uptake and metabolism by targeting CRL3 SPOP E3 ligase in cancer cells

    doi: 10.1038/s41467-022-30559-2

    Figure Lengend Snippet: a HEK293 cells were transfected with indicated plasmids, followed by immunoprecipitation with FLAG-agarose beads and immunoblotting. b HEK293 cells were transfected with various FLAG-CK1 isoforms, immunoprecipitated with FLAG-agarose beads and analyzed by immunoblotting. c Endogenous CK1δ protein in SUM159 cell lysates were pulled down with anti-S CK1δ antibody, and associated ASCT2 were detected by immunoblotting with anti-ASCT2 Ab (top, IP). Cell lysates were subjected to immunoblotting (bottom, input). d MDA-MB-231 cells were transfected with indicated CK1 isoforms, then analyzed by immunoblotting. e MDA-MB-231 cells were transfected siRNA targeting CK1δ for 48 h, followed by immunoblotting. f HEK293 cells were transfected with FLAG-SPOP and HA-ASCT2 and treated with D4476 for 12 h before harvesting, followed by immunoprecipitation with FLAG-agarose beads and analyzed by immunoblotting. g MDA-MB-231 cells were treated with different concentrations of CK1 inhibitor D4476 for 24 h, then analyzed by immunoblotting. h , i MDA-MB-231 cells were transfected with si-NC or siRNA targeting CK1δ (si-CK1δ#1), then incubated with CHX for various time points, and analyzed by immunoblotting ies ( h ), the band density of ASCT2 was quantified using ImageJ software and normalized to α-tubulin (mean ± SD, n = 3) ( i ). j HEK293 cells were transfected with indicated plasmids, lysed under denaturing conditions, followed by Ni-beads pulldown, and immunoblotting for ASCT2. Source data are provided as a Source Data file.

    Article Snippet: The following antibodies were used: ASCT2 (Cell Signaling Technology, D7C12, 8057, dilution: 1:1000); ASCT2 (Abcam, ab84903, dilution: 1:800); β-actin (Sigma-Aldrich, A5441, dilution: 1:10000); CK1δ (Santa Cruz, sc-55553, dilution: 1:1000); CK1δ (Cell Signaling Technology, 12417 S, dilution: 1:1000); CUL-1 (Santa Cruz, sc-11384, dilution: 1:1000); CUL-2 (Abcam, ab166917, dilution: 1:1000); CUL-3 (Cell Signaling Technology, 2759 S, dilution: 1:1000); CUL-4A (Cell Signaling Technology, 2699 S, dilution: 1:1000); CUL-4B (Proteintech, 12916-1-AP, dilution: 1:1000); CUL-5 (Santa Cruz, sc-13014, dilution: 1:1000); FLAG, clone M2 (Sigma-Aldrich, F1804-500UG, dilution: 1:2000); FLAG M2 affinity gel (Sigma-Aldrich, A2220-5ML); GLUD1 (Abcam, ab89967, dilution: 1:1000); GLS (Abcepta, ap8809b, dilution: 1:1000); GOT2 (ProteinTech, 14800-1-AP, dilution: 1:1000); GRK2 (ProteinTech, 13990-1-AP, dilution: 1:1000); GRK2 (Cell Signaling Technology, 3982, dilution: 1:1000); HA (Sigma, H6908, dilution: 1:2000); Anti-HA High Affinity (3F10) (Roche, 11867423001); HIF-1α (Santa Cruz, sc-53546, dilution: 1:1000); NAEβ (Abcam, ab124728, dilution: 1:1000); NAE1 (Cell Signaling Technology, 14321S, dilution: 1:1000); NEDD8 (Abcam, ab81264, dilution: 1:1000); SNAT1 (Abcam, ab134268, dilution: 1:1000); SNAT2 (Abcam, ab90677, dilution: 1:1000); SPOP (Abcam, ab137537, dilution: 1:1000); SPOP (Affinit, DF12106, dilution: 1:800); SPOP (Santa Cruz, sc-377206, dilution: 1:1000); p-SPOP (Ser222) (dilution: 1:1000); α-tubulin (Sigma, Clone AA13, T8203, dilution: 1:10000); UBE2M (Santa Cruz, sc-390064, dilution: 1:1000); UBE2F (ProteinTech, 17056-1-AP, dilution: 1:1000); Peroxidase AffiniPure Goat Anti-Rabbit IgG (H + L) (Jackson, 11-035-144, dilution: 1:4000); Peroxidase AffiniPure Goat Anti-Mouse IgG (H + L) (Jackson, 115-035-146, dilution: 1:4000); Peroxidase AffiniPure Goat Anti-Rat IgG (H + L) (Jackson, 112-035-143, dilution: 1:4000).

    Techniques: Transfection, Immunoprecipitation, Western Blot, Incubation, Software

    Cullin3 SPOP is the physiological E3 ubiquitin ligase for LATS1. (a) IB analysis of WCLs derived from 293T, 786-O and A498 cells treated with 10μM MG132 for 10 hr. (b) IB analysis of immunoprecipitates (IPs) and WCLs derived from 293T kidney cells transfected with indicated constructs. Cells were treated with MG132 (10 μM) before harvesting. (c) IB analysis of WCLs derived from A498 cells transfected with Cullin3 siRNA. (d) IB analysis of WCLs derived from 786-O cells after the specified duration of 100 μg/ml cycloheximide (CHX) transfected with Cullin3 siRNA. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated constructs. Cells were treated with MG132 (10μM) before harvesting. (g) IB analysis of WCLs derived from 293T kidney cells transfected with increasing doses of plasmid encoding SPOP. (h) IB analysis of WCLs derived from 786-O cells transfected with increasing doses of plasmid encoding SPOP. (i) IB analysis of WCLs derived from 293T kidney cells or A498 cells transfected with indicated constructs. Where indicated, cells were treated with 10μM MG132 before harvesting. (j) IB analysis of WCLs and products of ubiquitination derived from 293T cells transfected with indicated constructs.

    Journal: EBioMedicine

    Article Title: SPOP promotes ubiquitination and degradation of LATS1 to enhance kidney cancer progression

    doi: 10.1016/j.ebiom.2020.102795

    Figure Lengend Snippet: Cullin3 SPOP is the physiological E3 ubiquitin ligase for LATS1. (a) IB analysis of WCLs derived from 293T, 786-O and A498 cells treated with 10μM MG132 for 10 hr. (b) IB analysis of immunoprecipitates (IPs) and WCLs derived from 293T kidney cells transfected with indicated constructs. Cells were treated with MG132 (10 μM) before harvesting. (c) IB analysis of WCLs derived from A498 cells transfected with Cullin3 siRNA. (d) IB analysis of WCLs derived from 786-O cells after the specified duration of 100 μg/ml cycloheximide (CHX) transfected with Cullin3 siRNA. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated constructs. Cells were treated with MG132 (10μM) before harvesting. (g) IB analysis of WCLs derived from 293T kidney cells transfected with increasing doses of plasmid encoding SPOP. (h) IB analysis of WCLs derived from 786-O cells transfected with increasing doses of plasmid encoding SPOP. (i) IB analysis of WCLs derived from 293T kidney cells or A498 cells transfected with indicated constructs. Where indicated, cells were treated with 10μM MG132 before harvesting. (j) IB analysis of WCLs and products of ubiquitination derived from 293T cells transfected with indicated constructs.

    Article Snippet: Anti-Cul3(2759), anti-LATS1(3477) and anti-CK1(12417) antibodies were purchased from Cell Signaling.

    Techniques: Derivative Assay, Transfection, Construct, Plasmid Preparation

    Cullin3 SPOP E3 ubiquitin ligase negatively regulates the protein stability of LATS1. (a) IB analysis of WCLs derived from A498 or 786-O cells infected with the indicated lentiviral shRNA vectors. (b) IB analysis of WCLs derived from 293T or A498 cells transfected with HA-tagged SPOP plasmid. (c) Quantitative real-time PCR (qRT-PCR) analysis to detect LATS1 and SPOP mRNA levels derived from 786-O or A498 cells after depletion of SPOP. Data are shown as mean ±SD of three independent experiments. * P <0.05, Student's t test. (d) IB analysis of WCLs derived from 293T cells after the specified duration of cycloheximide (CHX) transfected with Myc-tagged SPOP plasmid. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs derived from A498 cells after the specified duration of 100μg/ml cycloheximide (CHX) infected with indicated lentiviral shRNA vectors. (g) The abundance of LATS1 protein in (f) was quantified and plotted.

    Journal: EBioMedicine

    Article Title: SPOP promotes ubiquitination and degradation of LATS1 to enhance kidney cancer progression

    doi: 10.1016/j.ebiom.2020.102795

    Figure Lengend Snippet: Cullin3 SPOP E3 ubiquitin ligase negatively regulates the protein stability of LATS1. (a) IB analysis of WCLs derived from A498 or 786-O cells infected with the indicated lentiviral shRNA vectors. (b) IB analysis of WCLs derived from 293T or A498 cells transfected with HA-tagged SPOP plasmid. (c) Quantitative real-time PCR (qRT-PCR) analysis to detect LATS1 and SPOP mRNA levels derived from 786-O or A498 cells after depletion of SPOP. Data are shown as mean ±SD of three independent experiments. * P <0.05, Student's t test. (d) IB analysis of WCLs derived from 293T cells after the specified duration of cycloheximide (CHX) transfected with Myc-tagged SPOP plasmid. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs derived from A498 cells after the specified duration of 100μg/ml cycloheximide (CHX) infected with indicated lentiviral shRNA vectors. (g) The abundance of LATS1 protein in (f) was quantified and plotted.

    Article Snippet: Anti-Cul3(2759), anti-LATS1(3477) and anti-CK1(12417) antibodies were purchased from Cell Signaling.

    Techniques: Derivative Assay, Infection, shRNA, Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    SPOP-mediated ubiquitination and degradation of LATS1 depends on the degron motif. (a) Amino acid sequence alignment of LATS1 with the SPOP–binding motif (degron) in known substrates of SPOP. (b) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (c) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting. (d) IB analysis of WCLs derived from 293T cells after the specified duration of 100μg/ml cycloheximide (CHX) transfected with indicated Flag-tagged LATS1 plasmids. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 hr before harvesting. (g) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting. (h) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (i) IB analysis of WCLs derived from 293T cells after the specified duration of 100μg/ml cycloheximide (CHX) transfected with indicated Flag-tagged LATS1 plasmids. (j) The abundance of LATS1 protein in (i) was quantified and plotted.

    Journal: EBioMedicine

    Article Title: SPOP promotes ubiquitination and degradation of LATS1 to enhance kidney cancer progression

    doi: 10.1016/j.ebiom.2020.102795

    Figure Lengend Snippet: SPOP-mediated ubiquitination and degradation of LATS1 depends on the degron motif. (a) Amino acid sequence alignment of LATS1 with the SPOP–binding motif (degron) in known substrates of SPOP. (b) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (c) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting. (d) IB analysis of WCLs derived from 293T cells after the specified duration of 100μg/ml cycloheximide (CHX) transfected with indicated Flag-tagged LATS1 plasmids. (e) The abundance of LATS1 protein in (d) was quantified and plotted. (f) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 hr before harvesting. (g) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting. (h) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (i) IB analysis of WCLs derived from 293T cells after the specified duration of 100μg/ml cycloheximide (CHX) transfected with indicated Flag-tagged LATS1 plasmids. (j) The abundance of LATS1 protein in (i) was quantified and plotted.

    Article Snippet: Anti-Cul3(2759), anti-LATS1(3477) and anti-CK1(12417) antibodies were purchased from Cell Signaling.

    Techniques: Sequencing, Binding Assay, Derivative Assay, Transfection

    CKΙδ promotes the interaction and degradation of LATS1 by SPOP. (a) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (b) IB analysis of WCLs derived from 293T, A498 and 786-O cells incubated with CKΙ inhibitor IC261 (50μM) or D4476 (20μM) before harvesting. (c) IB analysis of WCLs derived from kidney derived 293T cells transfected with CKΙδ siRNA. (d) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T kidney cells transfected with indicated plasmids and treated with 10μM MG132 for 10 hr before harvesting. (e) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (f) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (g) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting.

    Journal: EBioMedicine

    Article Title: SPOP promotes ubiquitination and degradation of LATS1 to enhance kidney cancer progression

    doi: 10.1016/j.ebiom.2020.102795

    Figure Lengend Snippet: CKΙδ promotes the interaction and degradation of LATS1 by SPOP. (a) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (b) IB analysis of WCLs derived from 293T, A498 and 786-O cells incubated with CKΙ inhibitor IC261 (50μM) or D4476 (20μM) before harvesting. (c) IB analysis of WCLs derived from kidney derived 293T cells transfected with CKΙδ siRNA. (d) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T kidney cells transfected with indicated plasmids and treated with 10μM MG132 for 10 hr before harvesting. (e) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (f) IB analysis of WCLs derived from 293T cells transfected with indicated plasmids. (g) IB analysis of WCLs and immunoprecipitates (IPs) derived from 293T cells transfected with indicated plasmids and treated with 10μM MG132 for 10 h before harvesting.

    Article Snippet: Anti-Cul3(2759), anti-LATS1(3477) and anti-CK1(12417) antibodies were purchased from Cell Signaling.

    Techniques: Derivative Assay, Transfection, Incubation

    SPOP promotes tumorigenesis. (a) 786-O-pcDNA3.1, 786-O-SPOP polyclonal stable kidney cancer cell lines were injected subcutaneously into the BALB/c-nu/nu mice. 40 days later, mice were anesthetic and taken a picture. (b) The tumours were dissected and taken a picture. (c) The weights of the dissected tumours in (b). (d) In vivo tumour growth was measured over the indicated time period. (e) The body weights of the BALB/c-nu/nu mice are measured over the indicated time period. (f) Immunohistochemistry staining of SPOP and LATS1 in tissue sections of xenografted tumours in BALB/c-nu/nu mice injected with the indicated cell lines. Scale bars, 20 μm. (g) IB analysis of the LATS1 protein levels in the dissected tumours. (h) Statistical analyses of the correlation between SPOP and LATS1 expression in cytoplasm of the kidney cancer tissue microarray. (i) Representative images of SPOP and LATS1 IHC from 89 cases of kidney cancer.

    Journal: EBioMedicine

    Article Title: SPOP promotes ubiquitination and degradation of LATS1 to enhance kidney cancer progression

    doi: 10.1016/j.ebiom.2020.102795

    Figure Lengend Snippet: SPOP promotes tumorigenesis. (a) 786-O-pcDNA3.1, 786-O-SPOP polyclonal stable kidney cancer cell lines were injected subcutaneously into the BALB/c-nu/nu mice. 40 days later, mice were anesthetic and taken a picture. (b) The tumours were dissected and taken a picture. (c) The weights of the dissected tumours in (b). (d) In vivo tumour growth was measured over the indicated time period. (e) The body weights of the BALB/c-nu/nu mice are measured over the indicated time period. (f) Immunohistochemistry staining of SPOP and LATS1 in tissue sections of xenografted tumours in BALB/c-nu/nu mice injected with the indicated cell lines. Scale bars, 20 μm. (g) IB analysis of the LATS1 protein levels in the dissected tumours. (h) Statistical analyses of the correlation between SPOP and LATS1 expression in cytoplasm of the kidney cancer tissue microarray. (i) Representative images of SPOP and LATS1 IHC from 89 cases of kidney cancer.

    Article Snippet: Anti-Cul3(2759), anti-LATS1(3477) and anti-CK1(12417) antibodies were purchased from Cell Signaling.

    Techniques: Injection, In Vivo, Immunohistochemistry, Staining, Expressing, Microarray