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p130 c 20 sc 317  (Santa Cruz Biotechnology)


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    Santa Cruz Biotechnology p130 c 20 sc 317
    P130 C 20 Sc 317, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 422 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p130+c+20+sc+317/p130/pm34716260-233-16-36
    Average 93 stars, based on 422 article reviews
    p130 c 20 sc 317 - by Bioz Stars, 2026-11
    93/100 stars

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    other:

    Article Title: MDM2, MDMX, and p73 regulate cell-cycle progression in the absence of wild-type p53.
    Article Snippet: Commercially obtained antibodies used in this study were as follows: pRB (IF-8) SC-102, p107 (C-18) SC-318, p130 (C-20) SC-317, E2F1 (KH95) SC-251, E2F2 (C-20) SC-318, E2F3 (C-18) SC-878, p21 (C-19) SC-397 (all SC are antibodies from Santa Cruz Biotechnology), p21 (12D1) (Cell Signaling Technology; 2947), p73 EP436Y (Abcam; ab40658), GAPDH 14C10 (Abcam; 2118), mouse HA-Tag (Cell Signaling Technology; C29F4), rabbit HA-Tag (BioLegend; 905102), actin (Sigma-Aldrich; A2066), and MDM2 DIV2Z (Cell Signaling Technology; 86934).



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    p130 c 20 sc 317  (Santa Cruz Biotechnology)
    93
    Santa Cruz Biotechnology p130 c 20 sc 317
    P130 C 20 Sc 317, supplied by Santa Cruz Biotechnology, 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/product/p130+c+20+sc+317/p130/pm34716260-233-16-36
    Average 93 stars, based on 1 article reviews
    p130 c 20 sc 317 - by Bioz Stars, 2026-11
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    p130 c 20 sc 317 affinity  (Santa Cruz Biotechnology)
    93
    Santa Cruz Biotechnology p130 c 20 sc 317 affinity
    Inhibition of MAGE-A11-dependent AR transcriptional activity by p107. A, pCMV5 (p5) or pCMV-AR (25 ng) was expressed in CV1 cells with 0.25 μg of PSA-Enh-Luc and 25 ng of empty vector (−), CMV-p107, or <t>pcDNA3-p130</t> with or without 50 ng of pSG5-MAGE (M) and 50 ng pSG5-HA-p300 (P). Cells were plated in medium containing 5% charcoal-stripped serum and incubated for 24 h the day after transfection with or without 1 nm DHT. B, LAPC-4 (LA, lane 1) and HeLa (HL, lane 2) cell extracts (80 μg of protein/lane) from cells cultured in serum-free medium for 24 h before harvest were prepared in immunoblot lysis buffer. Lane 3 contains 0.1 μg of protein extract from COS1 cells (C) expressing pSG5-MAGE. The immunoblot was probed using 10 μg/ml MAGE-A11 antibody. C, pCMV-AR (25 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pSG5 (−) or pSG5-MAGE with or without 50 ng of pSG5 (−) or 10, 25, or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. D, pCMV5 (25 ng) or 25 ng of pCMV-AR-(1–660) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc with or without 50 ng of pSG5-MAGE and pCMV5 (−), CMV-p107 and pSG5 (−), or pSG5-MAGE and CMV-p107. E, pCMV5 or pCMV-AR-(1–660) (10 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc, 50 ng of pSG5 (−) or pSG5-MAGE, and 50 ng of p5 (−) or 10, 25, or 50 ng of CMV-p107. F, pCMV5 (5 ng) (−), 5 ng of pCMV-AR-(1–660), or 25 ng of pCMV-AR was expressed in LAPC-4 cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pCMV5 (−) or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. In A and C–F, luciferase activity is the mean ± S.D. (error bars) and representative of three experiments.
    P130 C 20 Sc 317 Affinity, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    p130 c 20 sc 317 antibody  (Santa Cruz Biotechnology)
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    Santa Cruz Biotechnology p130 c 20 sc 317 antibody
    Inhibition of MAGE-A11-dependent AR transcriptional activity by p107. A, pCMV5 (p5) or pCMV-AR (25 ng) was expressed in CV1 cells with 0.25 μg of PSA-Enh-Luc and 25 ng of empty vector (−), CMV-p107, or <t>pcDNA3-p130</t> with or without 50 ng of pSG5-MAGE (M) and 50 ng pSG5-HA-p300 (P). Cells were plated in medium containing 5% charcoal-stripped serum and incubated for 24 h the day after transfection with or without 1 nm DHT. B, LAPC-4 (LA, lane 1) and HeLa (HL, lane 2) cell extracts (80 μg of protein/lane) from cells cultured in serum-free medium for 24 h before harvest were prepared in immunoblot lysis buffer. Lane 3 contains 0.1 μg of protein extract from COS1 cells (C) expressing pSG5-MAGE. The immunoblot was probed using 10 μg/ml MAGE-A11 antibody. C, pCMV-AR (25 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pSG5 (−) or pSG5-MAGE with or without 50 ng of pSG5 (−) or 10, 25, or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. D, pCMV5 (25 ng) or 25 ng of pCMV-AR-(1–660) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc with or without 50 ng of pSG5-MAGE and pCMV5 (−), CMV-p107 and pSG5 (−), or pSG5-MAGE and CMV-p107. E, pCMV5 or pCMV-AR-(1–660) (10 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc, 50 ng of pSG5 (−) or pSG5-MAGE, and 50 ng of p5 (−) or 10, 25, or 50 ng of CMV-p107. F, pCMV5 (5 ng) (−), 5 ng of pCMV-AR-(1–660), or 25 ng of pCMV-AR was expressed in LAPC-4 cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pCMV5 (−) or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. In A and C–F, luciferase activity is the mean ± S.D. (error bars) and representative of three experiments.
    P130 C 20 Sc 317 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit polyclonal anti p130 c 20 sc 317  (Santa Cruz Biotechnology)
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    Santa Cruz Biotechnology rabbit polyclonal anti p130 c 20 sc 317
    Inhibition of MAGE-A11-dependent AR transcriptional activity by p107. A, pCMV5 (p5) or pCMV-AR (25 ng) was expressed in CV1 cells with 0.25 μg of PSA-Enh-Luc and 25 ng of empty vector (−), CMV-p107, or <t>pcDNA3-p130</t> with or without 50 ng of pSG5-MAGE (M) and 50 ng pSG5-HA-p300 (P). Cells were plated in medium containing 5% charcoal-stripped serum and incubated for 24 h the day after transfection with or without 1 nm DHT. B, LAPC-4 (LA, lane 1) and HeLa (HL, lane 2) cell extracts (80 μg of protein/lane) from cells cultured in serum-free medium for 24 h before harvest were prepared in immunoblot lysis buffer. Lane 3 contains 0.1 μg of protein extract from COS1 cells (C) expressing pSG5-MAGE. The immunoblot was probed using 10 μg/ml MAGE-A11 antibody. C, pCMV-AR (25 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pSG5 (−) or pSG5-MAGE with or without 50 ng of pSG5 (−) or 10, 25, or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. D, pCMV5 (25 ng) or 25 ng of pCMV-AR-(1–660) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc with or without 50 ng of pSG5-MAGE and pCMV5 (−), CMV-p107 and pSG5 (−), or pSG5-MAGE and CMV-p107. E, pCMV5 or pCMV-AR-(1–660) (10 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc, 50 ng of pSG5 (−) or pSG5-MAGE, and 50 ng of p5 (−) or 10, 25, or 50 ng of CMV-p107. F, pCMV5 (5 ng) (−), 5 ng of pCMV-AR-(1–660), or 25 ng of pCMV-AR was expressed in LAPC-4 cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pCMV5 (−) or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. In A and C–F, luciferase activity is the mean ± S.D. (error bars) and representative of three experiments.
    Rabbit Polyclonal Anti P130 C 20 Sc 317, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit polyclonal anti p130 c 20 sc 317 - by Bioz Stars, 2026-11
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    2000 p130 c 20 sc  (Santa Cruz Biotechnology)
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    Santa Cruz Biotechnology 2000 p130 c 20 sc
    Inhibition of MAGE-A11-dependent AR transcriptional activity by p107. A, pCMV5 (p5) or pCMV-AR (25 ng) was expressed in CV1 cells with 0.25 μg of PSA-Enh-Luc and 25 ng of empty vector (−), CMV-p107, or <t>pcDNA3-p130</t> with or without 50 ng of pSG5-MAGE (M) and 50 ng pSG5-HA-p300 (P). Cells were plated in medium containing 5% charcoal-stripped serum and incubated for 24 h the day after transfection with or without 1 nm DHT. B, LAPC-4 (LA, lane 1) and HeLa (HL, lane 2) cell extracts (80 μg of protein/lane) from cells cultured in serum-free medium for 24 h before harvest were prepared in immunoblot lysis buffer. Lane 3 contains 0.1 μg of protein extract from COS1 cells (C) expressing pSG5-MAGE. The immunoblot was probed using 10 μg/ml MAGE-A11 antibody. C, pCMV-AR (25 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pSG5 (−) or pSG5-MAGE with or without 50 ng of pSG5 (−) or 10, 25, or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. D, pCMV5 (25 ng) or 25 ng of pCMV-AR-(1–660) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc with or without 50 ng of pSG5-MAGE and pCMV5 (−), CMV-p107 and pSG5 (−), or pSG5-MAGE and CMV-p107. E, pCMV5 or pCMV-AR-(1–660) (10 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc, 50 ng of pSG5 (−) or pSG5-MAGE, and 50 ng of p5 (−) or 10, 25, or 50 ng of CMV-p107. F, pCMV5 (5 ng) (−), 5 ng of pCMV-AR-(1–660), or 25 ng of pCMV-AR was expressed in LAPC-4 cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pCMV5 (−) or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. In A and C–F, luciferase activity is the mean ± S.D. (error bars) and representative of three experiments.
    2000 P130 C 20 Sc, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    p130 (c-20, sc-317) antibody  (Santa Cruz Biotechnology)
    90
    Santa Cruz Biotechnology p130 (c-20, sc-317) antibody
    Inhibition of MAGE-A11-dependent AR transcriptional activity by p107. A, pCMV5 (p5) or pCMV-AR (25 ng) was expressed in CV1 cells with 0.25 μg of PSA-Enh-Luc and 25 ng of empty vector (−), CMV-p107, or <t>pcDNA3-p130</t> with or without 50 ng of pSG5-MAGE (M) and 50 ng pSG5-HA-p300 (P). Cells were plated in medium containing 5% charcoal-stripped serum and incubated for 24 h the day after transfection with or without 1 nm DHT. B, LAPC-4 (LA, lane 1) and HeLa (HL, lane 2) cell extracts (80 μg of protein/lane) from cells cultured in serum-free medium for 24 h before harvest were prepared in immunoblot lysis buffer. Lane 3 contains 0.1 μg of protein extract from COS1 cells (C) expressing pSG5-MAGE. The immunoblot was probed using 10 μg/ml MAGE-A11 antibody. C, pCMV-AR (25 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pSG5 (−) or pSG5-MAGE with or without 50 ng of pSG5 (−) or 10, 25, or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. D, pCMV5 (25 ng) or 25 ng of pCMV-AR-(1–660) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc with or without 50 ng of pSG5-MAGE and pCMV5 (−), CMV-p107 and pSG5 (−), or pSG5-MAGE and CMV-p107. E, pCMV5 or pCMV-AR-(1–660) (10 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc, 50 ng of pSG5 (−) or pSG5-MAGE, and 50 ng of p5 (−) or 10, 25, or 50 ng of CMV-p107. F, pCMV5 (5 ng) (−), 5 ng of pCMV-AR-(1–660), or 25 ng of pCMV-AR was expressed in LAPC-4 cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pCMV5 (−) or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. In A and C–F, luciferase activity is the mean ± S.D. (error bars) and representative of three experiments.
    P130 (C 20, Sc 317) Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p130+c+20+sc+317/anti+p130/pmc02757496-48-19-6
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    p130 (c-20, sc-317) antibody - by Bioz Stars, 2026-11
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    p130 c 20 sc  (Santa Cruz Biotechnology)
    93
    Santa Cruz Biotechnology p130 c 20 sc
    Inhibition of MAGE-A11-dependent AR transcriptional activity by p107. A, pCMV5 (p5) or pCMV-AR (25 ng) was expressed in CV1 cells with 0.25 μg of PSA-Enh-Luc and 25 ng of empty vector (−), CMV-p107, or <t>pcDNA3-p130</t> with or without 50 ng of pSG5-MAGE (M) and 50 ng pSG5-HA-p300 (P). Cells were plated in medium containing 5% charcoal-stripped serum and incubated for 24 h the day after transfection with or without 1 nm DHT. B, LAPC-4 (LA, lane 1) and HeLa (HL, lane 2) cell extracts (80 μg of protein/lane) from cells cultured in serum-free medium for 24 h before harvest were prepared in immunoblot lysis buffer. Lane 3 contains 0.1 μg of protein extract from COS1 cells (C) expressing pSG5-MAGE. The immunoblot was probed using 10 μg/ml MAGE-A11 antibody. C, pCMV-AR (25 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pSG5 (−) or pSG5-MAGE with or without 50 ng of pSG5 (−) or 10, 25, or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. D, pCMV5 (25 ng) or 25 ng of pCMV-AR-(1–660) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc with or without 50 ng of pSG5-MAGE and pCMV5 (−), CMV-p107 and pSG5 (−), or pSG5-MAGE and CMV-p107. E, pCMV5 or pCMV-AR-(1–660) (10 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc, 50 ng of pSG5 (−) or pSG5-MAGE, and 50 ng of p5 (−) or 10, 25, or 50 ng of CMV-p107. F, pCMV5 (5 ng) (−), 5 ng of pCMV-AR-(1–660), or 25 ng of pCMV-AR was expressed in LAPC-4 cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pCMV5 (−) or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. In A and C–F, luciferase activity is the mean ± S.D. (error bars) and representative of three experiments.
    P130 C 20 Sc, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Inhibition of MAGE-A11-dependent AR transcriptional activity by p107. A, pCMV5 (p5) or pCMV-AR (25 ng) was expressed in CV1 cells with 0.25 μg of PSA-Enh-Luc and 25 ng of empty vector (−), CMV-p107, or pcDNA3-p130 with or without 50 ng of pSG5-MAGE (M) and 50 ng pSG5-HA-p300 (P). Cells were plated in medium containing 5% charcoal-stripped serum and incubated for 24 h the day after transfection with or without 1 nm DHT. B, LAPC-4 (LA, lane 1) and HeLa (HL, lane 2) cell extracts (80 μg of protein/lane) from cells cultured in serum-free medium for 24 h before harvest were prepared in immunoblot lysis buffer. Lane 3 contains 0.1 μg of protein extract from COS1 cells (C) expressing pSG5-MAGE. The immunoblot was probed using 10 μg/ml MAGE-A11 antibody. C, pCMV-AR (25 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pSG5 (−) or pSG5-MAGE with or without 50 ng of pSG5 (−) or 10, 25, or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. D, pCMV5 (25 ng) or 25 ng of pCMV-AR-(1–660) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc with or without 50 ng of pSG5-MAGE and pCMV5 (−), CMV-p107 and pSG5 (−), or pSG5-MAGE and CMV-p107. E, pCMV5 or pCMV-AR-(1–660) (10 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc, 50 ng of pSG5 (−) or pSG5-MAGE, and 50 ng of p5 (−) or 10, 25, or 50 ng of CMV-p107. F, pCMV5 (5 ng) (−), 5 ng of pCMV-AR-(1–660), or 25 ng of pCMV-AR was expressed in LAPC-4 cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pCMV5 (−) or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. In A and C–F, luciferase activity is the mean ± S.D. (error bars) and representative of three experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: Proto-oncogene Activity of Melanoma Antigen-A11 (MAGE-A11) Regulates Retinoblastoma-related p107 and E2F1 Proteins *

    doi: 10.1074/jbc.M113.468579

    Figure Lengend Snippet: Inhibition of MAGE-A11-dependent AR transcriptional activity by p107. A, pCMV5 (p5) or pCMV-AR (25 ng) was expressed in CV1 cells with 0.25 μg of PSA-Enh-Luc and 25 ng of empty vector (−), CMV-p107, or pcDNA3-p130 with or without 50 ng of pSG5-MAGE (M) and 50 ng pSG5-HA-p300 (P). Cells were plated in medium containing 5% charcoal-stripped serum and incubated for 24 h the day after transfection with or without 1 nm DHT. B, LAPC-4 (LA, lane 1) and HeLa (HL, lane 2) cell extracts (80 μg of protein/lane) from cells cultured in serum-free medium for 24 h before harvest were prepared in immunoblot lysis buffer. Lane 3 contains 0.1 μg of protein extract from COS1 cells (C) expressing pSG5-MAGE. The immunoblot was probed using 10 μg/ml MAGE-A11 antibody. C, pCMV-AR (25 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pSG5 (−) or pSG5-MAGE with or without 50 ng of pSG5 (−) or 10, 25, or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. D, pCMV5 (25 ng) or 25 ng of pCMV-AR-(1–660) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc with or without 50 ng of pSG5-MAGE and pCMV5 (−), CMV-p107 and pSG5 (−), or pSG5-MAGE and CMV-p107. E, pCMV5 or pCMV-AR-(1–660) (10 ng) was expressed in HeLa cells with 0.1 μg of PSA-Enh-Luc, 50 ng of pSG5 (−) or pSG5-MAGE, and 50 ng of p5 (−) or 10, 25, or 50 ng of CMV-p107. F, pCMV5 (5 ng) (−), 5 ng of pCMV-AR-(1–660), or 25 ng of pCMV-AR was expressed in LAPC-4 cells with 0.1 μg of PSA-Enh-Luc and 50 ng of pCMV5 (−) or 50 ng of CMV-p107. Cells were incubated with or without 1 nm DHT. In A and C–F, luciferase activity is the mean ± S.D. (error bars) and representative of three experiments.

    Article Snippet: Immunoblots were probed using the following antibodies from Santa Cruz Biotechnology, Inc. at 1:200 dilution unless indicated otherwise: p107 C-18 (sc-318) affinity-purified rabbit polyclonal antibody to a carboxyl-terminal human p107 peptide, E2F1 KH95 (sc-251) mouse monoclonal antibody to human E2F1 amino acid residues 342–386, E2F1 C-20 (sc-193) rabbit polyclonal antibody to a carboxyl-terminal epitope, E2F4 C-108 (sc-512) rabbit polyclonal antibody to human E2F4 amino acid residues 108–300, p130 C-20 (sc-317) affinity-purified rabbit polyclonal antibody to a human p130 carboxyl-terminal peptide, and DP-1 K-20 (sc-610) rabbit polyclonal antibody.

    Techniques: Inhibition, Activity Assay, Plasmid Preparation, Incubation, Transfection, Cell Culture, Western Blot, Lysis, Expressing, Luciferase

    MAGE-A11 interacts with p107 and Rb but not p130. pCMV-FLAG (−), pCMV-FLAG-MAGE, or FLAG-MAGE-(112–429) (5 μg) was expressed in COS1 cells with 5 μg of CMV-p107 (A), pCMV-hRb (B), or pcDNA3-p130 (C). Cells were incubated for 24 h in serum-free medium containing 0.1 μg/ml EGF and harvested in immunoprecipitation lysis buffer without deoxycholate or glycerol. Cell extracts (50 μg of protein/lane, right) and immunoprecipitates (IP, left) were probed using p107, Rb, p130, and MAGE-A11 antibodies. Arrows, p107, Rb, p130, FLAG-MAGE, and IgG.

    Journal: The Journal of Biological Chemistry

    Article Title: Proto-oncogene Activity of Melanoma Antigen-A11 (MAGE-A11) Regulates Retinoblastoma-related p107 and E2F1 Proteins *

    doi: 10.1074/jbc.M113.468579

    Figure Lengend Snippet: MAGE-A11 interacts with p107 and Rb but not p130. pCMV-FLAG (−), pCMV-FLAG-MAGE, or FLAG-MAGE-(112–429) (5 μg) was expressed in COS1 cells with 5 μg of CMV-p107 (A), pCMV-hRb (B), or pcDNA3-p130 (C). Cells were incubated for 24 h in serum-free medium containing 0.1 μg/ml EGF and harvested in immunoprecipitation lysis buffer without deoxycholate or glycerol. Cell extracts (50 μg of protein/lane, right) and immunoprecipitates (IP, left) were probed using p107, Rb, p130, and MAGE-A11 antibodies. Arrows, p107, Rb, p130, FLAG-MAGE, and IgG.

    Article Snippet: Immunoblots were probed using the following antibodies from Santa Cruz Biotechnology, Inc. at 1:200 dilution unless indicated otherwise: p107 C-18 (sc-318) affinity-purified rabbit polyclonal antibody to a carboxyl-terminal human p107 peptide, E2F1 KH95 (sc-251) mouse monoclonal antibody to human E2F1 amino acid residues 342–386, E2F1 C-20 (sc-193) rabbit polyclonal antibody to a carboxyl-terminal epitope, E2F4 C-108 (sc-512) rabbit polyclonal antibody to human E2F4 amino acid residues 108–300, p130 C-20 (sc-317) affinity-purified rabbit polyclonal antibody to a human p130 carboxyl-terminal peptide, and DP-1 K-20 (sc-610) rabbit polyclonal antibody.

    Techniques: Incubation, Immunoprecipitation, Lysis

    MAGE-A11 interacts with endogenous p107, stabilizes p107, and modulates p107 ubiquitination. A, pCMV5 or CMV-p107 (2 μg) was expressed in COS1 cells in 10-cm dishes with or without 2 μg of pSG5-MAGE. Cells were incubated for 24 h in serum-free medium with or without 0.1 μg/ml EGF. Cell extracts prepared in immunoblot lysis buffer (40 μg of protein/lane) were probed using p107 and MAGE-A11 antibodies. B, CMV-p107 (3 μg), 3 μg of pCMV5 (−), or 3 μg of pcDNA3-p130 was expressed in COS1 cells in 6-cm dishes with or without 1 μg of pSG5-MAGE. Cells were incubated for 24 h in serum-free medium with 0.1 μg/ml EGF. Cell extracts (40 μg of protein/lane) prepared in immunoblot lysis buffer were probed using p107, p130, and MAGE-A11 antibodies. C, pCMV5 (2 μg) alone (−) or 2 μg of CMV-p107 with or without 100 ng of WT pSG5-HA-MAGE-(2–429), 25 ng of pSG5-HA-MAGE-(112–429), 50 ng of pSG5-HA-MAGE-(112–307), or 100 ng of pSG5-HA-MAGE-(112–298) were expressed in COS1 cells in 10-cm dishes. Cells were incubated for 24 h in serum-free medium. Cell extracts (40 μg of protein/lane) prepared in immunoblot buffer were probed on transblots using p107 and HA antibodies. D, CMV-p107 (2 μg) and/or 2 μg of pSG5-MAGE were expressed in COS1 cells in 10-cm dishes. Cells were incubated for 24 h in serum-free medium. The next day, medium was exchanged, and cells were treated with 5 μm cycloheximide. Cells were harvested at 0, 8, 16, and 24 h in immunoblot lysis buffer. Cell extracts (50 μg of protein/lane) were probed using p107 and MAGE-A11 antibodies. E, pCMV-FLAG (6 μg) or 6 μg of pCMV-FLAG-ubiquitin (FLAG-Ub) was expressed in COS1 cells with or without 1 μg of pSG5-MAGE and/or 3 μg of CMV-p107. Cells were incubated for 24 h in serum-free medium containing 0.1 μg/ml EGF and immunoprecipitated using FLAG antibody affinity resin. Transblots of immunoprecipitates (IP, top panels) and cell extracts (40 μg of protein/lane, bottom panels) were probed using p107 and MAGE-A11 antibodies.

    Journal: The Journal of Biological Chemistry

    Article Title: Proto-oncogene Activity of Melanoma Antigen-A11 (MAGE-A11) Regulates Retinoblastoma-related p107 and E2F1 Proteins *

    doi: 10.1074/jbc.M113.468579

    Figure Lengend Snippet: MAGE-A11 interacts with endogenous p107, stabilizes p107, and modulates p107 ubiquitination. A, pCMV5 or CMV-p107 (2 μg) was expressed in COS1 cells in 10-cm dishes with or without 2 μg of pSG5-MAGE. Cells were incubated for 24 h in serum-free medium with or without 0.1 μg/ml EGF. Cell extracts prepared in immunoblot lysis buffer (40 μg of protein/lane) were probed using p107 and MAGE-A11 antibodies. B, CMV-p107 (3 μg), 3 μg of pCMV5 (−), or 3 μg of pcDNA3-p130 was expressed in COS1 cells in 6-cm dishes with or without 1 μg of pSG5-MAGE. Cells were incubated for 24 h in serum-free medium with 0.1 μg/ml EGF. Cell extracts (40 μg of protein/lane) prepared in immunoblot lysis buffer were probed using p107, p130, and MAGE-A11 antibodies. C, pCMV5 (2 μg) alone (−) or 2 μg of CMV-p107 with or without 100 ng of WT pSG5-HA-MAGE-(2–429), 25 ng of pSG5-HA-MAGE-(112–429), 50 ng of pSG5-HA-MAGE-(112–307), or 100 ng of pSG5-HA-MAGE-(112–298) were expressed in COS1 cells in 10-cm dishes. Cells were incubated for 24 h in serum-free medium. Cell extracts (40 μg of protein/lane) prepared in immunoblot buffer were probed on transblots using p107 and HA antibodies. D, CMV-p107 (2 μg) and/or 2 μg of pSG5-MAGE were expressed in COS1 cells in 10-cm dishes. Cells were incubated for 24 h in serum-free medium. The next day, medium was exchanged, and cells were treated with 5 μm cycloheximide. Cells were harvested at 0, 8, 16, and 24 h in immunoblot lysis buffer. Cell extracts (50 μg of protein/lane) were probed using p107 and MAGE-A11 antibodies. E, pCMV-FLAG (6 μg) or 6 μg of pCMV-FLAG-ubiquitin (FLAG-Ub) was expressed in COS1 cells with or without 1 μg of pSG5-MAGE and/or 3 μg of CMV-p107. Cells were incubated for 24 h in serum-free medium containing 0.1 μg/ml EGF and immunoprecipitated using FLAG antibody affinity resin. Transblots of immunoprecipitates (IP, top panels) and cell extracts (40 μg of protein/lane, bottom panels) were probed using p107 and MAGE-A11 antibodies.

    Article Snippet: Immunoblots were probed using the following antibodies from Santa Cruz Biotechnology, Inc. at 1:200 dilution unless indicated otherwise: p107 C-18 (sc-318) affinity-purified rabbit polyclonal antibody to a carboxyl-terminal human p107 peptide, E2F1 KH95 (sc-251) mouse monoclonal antibody to human E2F1 amino acid residues 342–386, E2F1 C-20 (sc-193) rabbit polyclonal antibody to a carboxyl-terminal epitope, E2F4 C-108 (sc-512) rabbit polyclonal antibody to human E2F4 amino acid residues 108–300, p130 C-20 (sc-317) affinity-purified rabbit polyclonal antibody to a human p130 carboxyl-terminal peptide, and DP-1 K-20 (sc-610) rabbit polyclonal antibody.

    Techniques: Incubation, Western Blot, Lysis, Immunoprecipitation

    p107 inhibits MAGE-A11 activation of E2F1 in HeLa cells. A, pSG5 (25 ng) (−) or 25 ng of pSG5-MAGE was expressed in HeLa cells with 0.1 μg of E2F1-Luc with or without 50 ng of pSG5 (−) or pSG5-HA-p300 and 10 ng of pCMV5 (−) or CMV-p107. B, pSG5 (50 ng) alone (−), 25 ng of pSG5-MAGE (left), or 25 ng of CMX-E1A (right) was expressed in HeLa cells with 0.1 μg of E2F1-Luc with 50 ng of pCMV5 (−) or 10, 25, or 50 ng of CMV-p107. C, pCMV5 (100 ng) (−) or 100 ng of pSG5-MAGE was expressed in HeLa cells with 0.1 μg of E2F1-Luc and 10 ng of pCMV5 (−), CMV-p107, CMV-p107Δ409–826 (DE) or pcDNA3-p130. Luciferase activity is the mean ± S.D. (error bars) representative of three experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: Proto-oncogene Activity of Melanoma Antigen-A11 (MAGE-A11) Regulates Retinoblastoma-related p107 and E2F1 Proteins *

    doi: 10.1074/jbc.M113.468579

    Figure Lengend Snippet: p107 inhibits MAGE-A11 activation of E2F1 in HeLa cells. A, pSG5 (25 ng) (−) or 25 ng of pSG5-MAGE was expressed in HeLa cells with 0.1 μg of E2F1-Luc with or without 50 ng of pSG5 (−) or pSG5-HA-p300 and 10 ng of pCMV5 (−) or CMV-p107. B, pSG5 (50 ng) alone (−), 25 ng of pSG5-MAGE (left), or 25 ng of CMX-E1A (right) was expressed in HeLa cells with 0.1 μg of E2F1-Luc with 50 ng of pCMV5 (−) or 10, 25, or 50 ng of CMV-p107. C, pCMV5 (100 ng) (−) or 100 ng of pSG5-MAGE was expressed in HeLa cells with 0.1 μg of E2F1-Luc and 10 ng of pCMV5 (−), CMV-p107, CMV-p107Δ409–826 (DE) or pcDNA3-p130. Luciferase activity is the mean ± S.D. (error bars) representative of three experiments.

    Article Snippet: Immunoblots were probed using the following antibodies from Santa Cruz Biotechnology, Inc. at 1:200 dilution unless indicated otherwise: p107 C-18 (sc-318) affinity-purified rabbit polyclonal antibody to a carboxyl-terminal human p107 peptide, E2F1 KH95 (sc-251) mouse monoclonal antibody to human E2F1 amino acid residues 342–386, E2F1 C-20 (sc-193) rabbit polyclonal antibody to a carboxyl-terminal epitope, E2F4 C-108 (sc-512) rabbit polyclonal antibody to human E2F4 amino acid residues 108–300, p130 C-20 (sc-317) affinity-purified rabbit polyclonal antibody to a human p130 carboxyl-terminal peptide, and DP-1 K-20 (sc-610) rabbit polyclonal antibody.

    Techniques: Activation Assay, Luciferase, Activity Assay