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    Cell Signaling Technology Inc mean sd 64 4 14 2 65 5
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    mean sd 64 4 14 2 65 5  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc mean sd 64 4 14 2 65 5
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    mean sd 14 0 57 6 40 9  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc cell surface receptors
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    Cell Signaling Technology Inc mouse ps6
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    Cell Signaling Technology Inc wnt5a 3p21 p14 cell signaling 65
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    anti phospho serine 65 4e bp1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho serine 65 4e bp1
    (A.) T47D cells were treated for 24 hours with 10μM antagonists (IPAG, haloperidol) or 10μM agonists (PRE084, (+)SKF-10047). Subsequently, cells were pulse-labeled for 1 hour with 100μCi/ml [35S] methionine and cysteine. Protein extracts were resolved by SDS-PAGE, transferred onto nitrocellulose membrane and exposed to autoradiography film. (B.) Immunoblots of protein extracts from T47D cells treated for 24 hours with 10μM putative antagonists (IPAG, haloperidol) or putative agonists (PRE084, (+)SKF-10047). Phospho-threonine 389-p70S6K (P-S6K), phospho-serine 235/236 S6 (P-S6), and <t>phospho-serine</t> <t>65-4E-BP1</t> (P-4E-BP1). (C.) Cell lysates were precipitated with m7GTP-sepharose beads (pull-down) and subsequently immunoblotted with antibodies against 4E-BP1 and eIF4E to evaluate 4E-BP1 binding to the eIF4E-mRNA cap complex. The upper panel (cell lysates) demonstrates equivalent input into m7GTP-sepharose binding, as well as Sigma1 ligand mediated changes in 4E-BP1 phosphorylation profile. (D.) Immunoblots of protein extracts from breast (MDA468, MCF7) and prostate adenocarcinoma (PC3, LNCaP) cells treated for 24 hours with 10μM IPAG.
    Anti Phospho Serine 65 4e Bp1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Inhibition of tumor cell growth by Sigma1 ligand mediated translational repression"

    Article Title: Inhibition of tumor cell growth by Sigma1 ligand mediated translational repression

    Journal: Biochemical and biophysical research communications

    doi: 10.1016/j.bbrc.2012.08.052

    (A.) T47D cells were treated for 24 hours with 10μM antagonists (IPAG, haloperidol) or 10μM agonists (PRE084, (+)SKF-10047). Subsequently, cells were pulse-labeled for 1 hour with 100μCi/ml [35S] methionine and cysteine. Protein extracts were resolved by SDS-PAGE, transferred onto nitrocellulose membrane and exposed to autoradiography film. (B.) Immunoblots of protein extracts from T47D cells treated for 24 hours with 10μM putative antagonists (IPAG, haloperidol) or putative agonists (PRE084, (+)SKF-10047). Phospho-threonine 389-p70S6K (P-S6K), phospho-serine 235/236 S6 (P-S6), and phospho-serine 65-4E-BP1 (P-4E-BP1). (C.) Cell lysates were precipitated with m7GTP-sepharose beads (pull-down) and subsequently immunoblotted with antibodies against 4E-BP1 and eIF4E to evaluate 4E-BP1 binding to the eIF4E-mRNA cap complex. The upper panel (cell lysates) demonstrates equivalent input into m7GTP-sepharose binding, as well as Sigma1 ligand mediated changes in 4E-BP1 phosphorylation profile. (D.) Immunoblots of protein extracts from breast (MDA468, MCF7) and prostate adenocarcinoma (PC3, LNCaP) cells treated for 24 hours with 10μM IPAG.
    Figure Legend Snippet: (A.) T47D cells were treated for 24 hours with 10μM antagonists (IPAG, haloperidol) or 10μM agonists (PRE084, (+)SKF-10047). Subsequently, cells were pulse-labeled for 1 hour with 100μCi/ml [35S] methionine and cysteine. Protein extracts were resolved by SDS-PAGE, transferred onto nitrocellulose membrane and exposed to autoradiography film. (B.) Immunoblots of protein extracts from T47D cells treated for 24 hours with 10μM putative antagonists (IPAG, haloperidol) or putative agonists (PRE084, (+)SKF-10047). Phospho-threonine 389-p70S6K (P-S6K), phospho-serine 235/236 S6 (P-S6), and phospho-serine 65-4E-BP1 (P-4E-BP1). (C.) Cell lysates were precipitated with m7GTP-sepharose beads (pull-down) and subsequently immunoblotted with antibodies against 4E-BP1 and eIF4E to evaluate 4E-BP1 binding to the eIF4E-mRNA cap complex. The upper panel (cell lysates) demonstrates equivalent input into m7GTP-sepharose binding, as well as Sigma1 ligand mediated changes in 4E-BP1 phosphorylation profile. (D.) Immunoblots of protein extracts from breast (MDA468, MCF7) and prostate adenocarcinoma (PC3, LNCaP) cells treated for 24 hours with 10μM IPAG.

    Techniques Used: Labeling, SDS Page, Autoradiography, Western Blot, Binding Assay

    (A.) Approximately 4.5 days post-transfection, translational control markers phospho-threonine 389-p70S6K (P-S6K), phospho-serine 235/236 S6 (P-S6), and phospho-serine 65-4E-BP1 (P-4E-BP1) were evaluated in immunoblotted T47D protein extracts. (B.) Following transfection of control or Sigma1 siRNA, T47D cells were treated for 24 hours with 10μM IPAG and the translational control markers, above, were detected by immunoblot.
    Figure Legend Snippet: (A.) Approximately 4.5 days post-transfection, translational control markers phospho-threonine 389-p70S6K (P-S6K), phospho-serine 235/236 S6 (P-S6), and phospho-serine 65-4E-BP1 (P-4E-BP1) were evaluated in immunoblotted T47D protein extracts. (B.) Following transfection of control or Sigma1 siRNA, T47D cells were treated for 24 hours with 10μM IPAG and the translational control markers, above, were detected by immunoblot.

    Techniques Used: Transfection, Western Blot

    T47D cells were treated with 10μM IPAG, and cell culture medium was changed with drug-free medium after treatment for 24 hours. Cells in “recovery” were treated as follows: cells were immediately washed with pre-warmed culture medium to remove residual drug (washed out). Recovery indicates T47D cell cultures in which drug-containing medium was removed and replaced with drug-free medium after 24 hour treatment with 10μM of IPAG (antagonist). Cells were collected and analyzed 24, 48, and 72 hours after removal of drug from cell culture medium FSC-H was measured at the indicated treatment time. (A.) T47D cells were either continuously treated with 10μM of IPAG (antagonist) for 96 hours (open square, solid line) and compared to DMSO treated cells (closed circle, solid line), or compared to cells that were treated with 10μM of IPAG (antagonist) for 24 hours and subsequently allowed to recover following removal of IPAG from cell culture medium (open triangle, broken line) (n=3, **P<0.01). The FSC-H of cells at the 48 hour recovery time point are not significantly different compared to control cells that have been treated for 72 hours with DMSO. Error bars are not shown since they are smaller than the size of the symbols. (B.) Cell death was monitored by Trypan blue exclusion assay following treatment with 10μM IPAG for the indicated times (n=3, **P<0.01). (C.) Restored protein synthesis 24 and 48 hours following IPAG wash-out. Protein extracted IPAG (10μM) treated T47D cells and evaluated for cell death in Figure 4B were resolved by SDS-PAGE and immunoblotted with antibodies against phospho-threonine 389-p70S6K (P-S6K), phospho-serine 235/236-S6 (P-S6), and phospho-serine 65-4E-BP1 (P-4EBP1). (D.) Suppressed protein synthesis following 48 hours of continuous treatment with 10μM IPAG.
    Figure Legend Snippet: T47D cells were treated with 10μM IPAG, and cell culture medium was changed with drug-free medium after treatment for 24 hours. Cells in “recovery” were treated as follows: cells were immediately washed with pre-warmed culture medium to remove residual drug (washed out). Recovery indicates T47D cell cultures in which drug-containing medium was removed and replaced with drug-free medium after 24 hour treatment with 10μM of IPAG (antagonist). Cells were collected and analyzed 24, 48, and 72 hours after removal of drug from cell culture medium FSC-H was measured at the indicated treatment time. (A.) T47D cells were either continuously treated with 10μM of IPAG (antagonist) for 96 hours (open square, solid line) and compared to DMSO treated cells (closed circle, solid line), or compared to cells that were treated with 10μM of IPAG (antagonist) for 24 hours and subsequently allowed to recover following removal of IPAG from cell culture medium (open triangle, broken line) (n=3, **P<0.01). The FSC-H of cells at the 48 hour recovery time point are not significantly different compared to control cells that have been treated for 72 hours with DMSO. Error bars are not shown since they are smaller than the size of the symbols. (B.) Cell death was monitored by Trypan blue exclusion assay following treatment with 10μM IPAG for the indicated times (n=3, **P<0.01). (C.) Restored protein synthesis 24 and 48 hours following IPAG wash-out. Protein extracted IPAG (10μM) treated T47D cells and evaluated for cell death in Figure 4B were resolved by SDS-PAGE and immunoblotted with antibodies against phospho-threonine 389-p70S6K (P-S6K), phospho-serine 235/236-S6 (P-S6), and phospho-serine 65-4E-BP1 (P-4EBP1). (D.) Suppressed protein synthesis following 48 hours of continuous treatment with 10μM IPAG.

    Techniques Used: Cell Culture, Trypan Blue Exclusion Assay, SDS Page

    surface receptors ecm up  (Cell Signaling Technology Inc)


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