rapamycin Search Results


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  • 99
    Millipore rapamycin sirolimus
    <t>Rapamycin</t> inhibits the spatial interaction of TLR9 with the MyD88-IRF7 complex and affects the phosphorylation and nuclear translocation of IRF7. ( a ) Immunoassay of RAW cells (4 × 10 6 ) transfected with hemagglutinin-tagged MyD88 and YFP-tagged TLR9 and then, 36 h later, stimulated for 90 min with CpG-A–DOTAP; cell extracts were immunoprecipitated (IP) with anti-hemagglutinin and analyzed by immunoblot (IB) for coprecipitated TLR9. MyD88 serves as a loading control. ( b ) Flow cytometry of human pDCs pretreated with rapamycin (left) or transfected with control (con) siRNA or siRNA specific for p70S6K1 and p70S6K2 (S6K), then stimulated for 45 min with CpG-A; cells were fixed, made permeable and stained with phycoerythrin-conjugated mouse antibody to IRF7 phosphorylated at Ser477 and Ser479 (phosphor-IRF7) or control isotype antibody (Control Ab; phycoerythrin-conjugated mouse immunoglobulin G1). ( c ) Confocal microscopy of purified mouse pDCs stimulated for 12 h with CpG-A in the presence of rapamycin or vehicle, then fixed with formaldehyde, made permeable with saponin, blocked with 20% (vol/vol) goat serum, stained with anti-IRF7 (green) and mounted with ProLong Gold antifade reagent with DAPI (blue). Scale bar, 2 μm. ( d ) Luciferase assay of HEK293 cells transiently transfected with 0 ng (−), 40 ng or 200 ng of constitutively active IRF7 (IRF7-D477,479) together with luciferase-tagged IFN-β (plasmid p125-Luc; 50 ng), renilla luciferase (0.5 ng), wild-type IRF7 (3 ng), MyD88 (20 ng) and TLR9 (50 ng), cultured for 24 h, then pretreated for 3 h with 100 nM rapamycin and then stimulated overnight with CpG-A (10 μg/ml). Empty vector, vector without IRF7. Data are representative of three ( a , d ) or two ( b , c ) independent experiments (error bars ( d ), s.d.).
    Rapamycin Sirolimus, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rapamycin sirolimus/product/Millipore
    Average 99 stars, based on 8 article reviews
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    85
    Wyeth Biopharma sirolimus rapamune
    Blood concentrations and biliary excretion of (A,C) <t>sirolimus</t> and (B,D) tacrolimus after daily treatment over 2 weeks with sirolimus (SRL, 1 mg kg −1 p.o.), tacrolimus (TRL 1 mg kg −1 i.p.) and of SRL (1 mg kg −1 p.o.) plus CyA (10 mg kg −1 i.p.) or TRL (1 mg kg −1 i.p.). Means±s.e.mean of 10 animals each. * Significant difference ( P
    Sirolimus Rapamune, supplied by Wyeth Biopharma, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirolimus rapamune/product/Wyeth Biopharma
    Average 85 stars, based on 3 article reviews
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    85
    LC Laboratories sirolimus rapamune
    TS-1/22 plus basiliximab and <t>sirolimus</t> prolongs islet allograft survival.
    Sirolimus Rapamune, supplied by LC Laboratories, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirolimus rapamune/product/LC Laboratories
    Average 85 stars, based on 8 article reviews
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    99
    Selleck Chemicals rapamycin sirolimus
    MoSNT2 is associated with the MoTor signaling pathway. (A) Vegetative growth of M. oryzae on CM agar medium supplemented with or without 1 μg/ml <t>rapamycin</t> (rapa.). (B) Inhibition rate of rapamycin on the mycelial growth. (C) Expression profiles of MoSNT2 and MoTOR in the wild-type Guy11 strain at different developmental processes. (D) Linear correlation between qRT-PCR-measured expression levels of MoSNT2 and MoTOR . (E) qRT-PCR analysis of MoSNT2 expression levels in the Guy11 strain in response to rapamycin. The Guy11 strain grown in liquid CM for 48 h was transferred into fresh liquid CM in the presence or absence of 1 μg/ml rapamycin for 6 h before total RNA extraction.
    Rapamycin Sirolimus, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 99/100, based on 117 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rapamycin sirolimus/product/Selleck Chemicals
    Average 99 stars, based on 117 article reviews
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    rapamycin sirolimus - by Bioz Stars, 2020-05
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    90
    Pfizer Inc sirolimus rapamune pfizer
    MoSNT2 is associated with the MoTor signaling pathway. (A) Vegetative growth of M. oryzae on CM agar medium supplemented with or without 1 μg/ml <t>rapamycin</t> (rapa.). (B) Inhibition rate of rapamycin on the mycelial growth. (C) Expression profiles of MoSNT2 and MoTOR in the wild-type Guy11 strain at different developmental processes. (D) Linear correlation between qRT-PCR-measured expression levels of MoSNT2 and MoTOR . (E) qRT-PCR analysis of MoSNT2 expression levels in the Guy11 strain in response to rapamycin. The Guy11 strain grown in liquid CM for 48 h was transferred into fresh liquid CM in the presence or absence of 1 μg/ml rapamycin for 6 h before total RNA extraction.
    Sirolimus Rapamune Pfizer, supplied by Pfizer Inc, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirolimus rapamune pfizer/product/Pfizer Inc
    Average 90 stars, based on 2 article reviews
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    93
    Wyeth Biopharma rapamune sirolimus immunosuppressant
    MoSNT2 is associated with the MoTor signaling pathway. (A) Vegetative growth of M. oryzae on CM agar medium supplemented with or without 1 μg/ml <t>rapamycin</t> (rapa.). (B) Inhibition rate of rapamycin on the mycelial growth. (C) Expression profiles of MoSNT2 and MoTOR in the wild-type Guy11 strain at different developmental processes. (D) Linear correlation between qRT-PCR-measured expression levels of MoSNT2 and MoTOR . (E) qRT-PCR analysis of MoSNT2 expression levels in the Guy11 strain in response to rapamycin. The Guy11 strain grown in liquid CM for 48 h was transferred into fresh liquid CM in the presence or absence of 1 μg/ml rapamycin for 6 h before total RNA extraction.
    Rapamune Sirolimus Immunosuppressant, supplied by Wyeth Biopharma, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rapamune sirolimus immunosuppressant/product/Wyeth Biopharma
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    Image Search Results


    Rapamycin inhibits the spatial interaction of TLR9 with the MyD88-IRF7 complex and affects the phosphorylation and nuclear translocation of IRF7. ( a ) Immunoassay of RAW cells (4 × 10 6 ) transfected with hemagglutinin-tagged MyD88 and YFP-tagged TLR9 and then, 36 h later, stimulated for 90 min with CpG-A–DOTAP; cell extracts were immunoprecipitated (IP) with anti-hemagglutinin and analyzed by immunoblot (IB) for coprecipitated TLR9. MyD88 serves as a loading control. ( b ) Flow cytometry of human pDCs pretreated with rapamycin (left) or transfected with control (con) siRNA or siRNA specific for p70S6K1 and p70S6K2 (S6K), then stimulated for 45 min with CpG-A; cells were fixed, made permeable and stained with phycoerythrin-conjugated mouse antibody to IRF7 phosphorylated at Ser477 and Ser479 (phosphor-IRF7) or control isotype antibody (Control Ab; phycoerythrin-conjugated mouse immunoglobulin G1). ( c ) Confocal microscopy of purified mouse pDCs stimulated for 12 h with CpG-A in the presence of rapamycin or vehicle, then fixed with formaldehyde, made permeable with saponin, blocked with 20% (vol/vol) goat serum, stained with anti-IRF7 (green) and mounted with ProLong Gold antifade reagent with DAPI (blue). Scale bar, 2 μm. ( d ) Luciferase assay of HEK293 cells transiently transfected with 0 ng (−), 40 ng or 200 ng of constitutively active IRF7 (IRF7-D477,479) together with luciferase-tagged IFN-β (plasmid p125-Luc; 50 ng), renilla luciferase (0.5 ng), wild-type IRF7 (3 ng), MyD88 (20 ng) and TLR9 (50 ng), cultured for 24 h, then pretreated for 3 h with 100 nM rapamycin and then stimulated overnight with CpG-A (10 μg/ml). Empty vector, vector without IRF7. Data are representative of three ( a , d ) or two ( b , c ) independent experiments (error bars ( d ), s.d.).

    Journal: Nature immunology

    Article Title: Toll-like receptor-mediated induction of type I interferon in plasmacytoid dendritic cells requires the rapamycin-sensitive PI(3)K-mTOR-p70S6K pathway

    doi: 10.1038/ni.1645

    Figure Lengend Snippet: Rapamycin inhibits the spatial interaction of TLR9 with the MyD88-IRF7 complex and affects the phosphorylation and nuclear translocation of IRF7. ( a ) Immunoassay of RAW cells (4 × 10 6 ) transfected with hemagglutinin-tagged MyD88 and YFP-tagged TLR9 and then, 36 h later, stimulated for 90 min with CpG-A–DOTAP; cell extracts were immunoprecipitated (IP) with anti-hemagglutinin and analyzed by immunoblot (IB) for coprecipitated TLR9. MyD88 serves as a loading control. ( b ) Flow cytometry of human pDCs pretreated with rapamycin (left) or transfected with control (con) siRNA or siRNA specific for p70S6K1 and p70S6K2 (S6K), then stimulated for 45 min with CpG-A; cells were fixed, made permeable and stained with phycoerythrin-conjugated mouse antibody to IRF7 phosphorylated at Ser477 and Ser479 (phosphor-IRF7) or control isotype antibody (Control Ab; phycoerythrin-conjugated mouse immunoglobulin G1). ( c ) Confocal microscopy of purified mouse pDCs stimulated for 12 h with CpG-A in the presence of rapamycin or vehicle, then fixed with formaldehyde, made permeable with saponin, blocked with 20% (vol/vol) goat serum, stained with anti-IRF7 (green) and mounted with ProLong Gold antifade reagent with DAPI (blue). Scale bar, 2 μm. ( d ) Luciferase assay of HEK293 cells transiently transfected with 0 ng (−), 40 ng or 200 ng of constitutively active IRF7 (IRF7-D477,479) together with luciferase-tagged IFN-β (plasmid p125-Luc; 50 ng), renilla luciferase (0.5 ng), wild-type IRF7 (3 ng), MyD88 (20 ng) and TLR9 (50 ng), cultured for 24 h, then pretreated for 3 h with 100 nM rapamycin and then stimulated overnight with CpG-A (10 μg/ml). Empty vector, vector without IRF7. Data are representative of three ( a , d ) or two ( b , c ) independent experiments (error bars ( d ), s.d.).

    Article Snippet: CsA, FK506 (tacrolimus) and rapamycin (sirolimus) were from Sigma.

    Techniques: Translocation Assay, Transfection, Immunoprecipitation, Flow Cytometry, Cytometry, Staining, Confocal Microscopy, Purification, Luciferase, Plasmid Preparation, Cell Culture

    Inhibition of mTOR in APCs in vivo can suppress the adaptive immune response to a vaccine. IFN-γ production (top right) and cytokine expression (top left and bottom) by CD8 + T cells enriched from draining lymph nodes of C57BL/6 mice left untreated (−) or treated daily for 3 d with vehicle or rapamycin microparticles (2 mg per mouse per day) and then vaccinated subcutaneously on day 4 with YF-17D; 5 d later, single-cell suspensions were cultured for an additional 3 d in vitro in medium alone (left bars) or with a CD8 + T cell epitope from YF-17D (middle and right bars). IFN-γ was assessed by intracellular cytokine staining and flow cytometry; numbers above outlined areas indicate percent CD8 + IFN-γ + cells. IFN-γ, IL-4, IL-13, IL-10 and IL-17 in cell supernatants were measured by ELISA. *, P

    Journal: Nature immunology

    Article Title: Toll-like receptor-mediated induction of type I interferon in plasmacytoid dendritic cells requires the rapamycin-sensitive PI(3)K-mTOR-p70S6K pathway

    doi: 10.1038/ni.1645

    Figure Lengend Snippet: Inhibition of mTOR in APCs in vivo can suppress the adaptive immune response to a vaccine. IFN-γ production (top right) and cytokine expression (top left and bottom) by CD8 + T cells enriched from draining lymph nodes of C57BL/6 mice left untreated (−) or treated daily for 3 d with vehicle or rapamycin microparticles (2 mg per mouse per day) and then vaccinated subcutaneously on day 4 with YF-17D; 5 d later, single-cell suspensions were cultured for an additional 3 d in vitro in medium alone (left bars) or with a CD8 + T cell epitope from YF-17D (middle and right bars). IFN-γ was assessed by intracellular cytokine staining and flow cytometry; numbers above outlined areas indicate percent CD8 + IFN-γ + cells. IFN-γ, IL-4, IL-13, IL-10 and IL-17 in cell supernatants were measured by ELISA. *, P

    Article Snippet: CsA, FK506 (tacrolimus) and rapamycin (sirolimus) were from Sigma.

    Techniques: Inhibition, In Vivo, Expressing, Mouse Assay, Cell Culture, In Vitro, Staining, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    Expression of rapamycin-sensitive mTOR pathway components in pDCs. ( a ) Immunoblot analysis of phosphorylated (phospho-) and total mTOR in lysates of RAW cells (4 × 10 6 ) transiently transfected for 40 h with cyan fluorescent protein–tagged MyD88 and IRF7-YFP, treated with vehicle (−) or rapamycin (+) and then stimulated for 0–60 min with CpG-A–DOTAP. Results are representative of two independent experiments. ( b ) Immunoblot analysis of phosphorylated and total mTOR, p70S6K, 4E-BP1 and Akt in lysates of purified pDCs (1 × 10 6 ) stimulated for 15 min with CpG-A in the presence (Rap) or absence (Medium) of rapamycin. Results are representative of three independent experiments.

    Journal: Nature immunology

    Article Title: Toll-like receptor-mediated induction of type I interferon in plasmacytoid dendritic cells requires the rapamycin-sensitive PI(3)K-mTOR-p70S6K pathway

    doi: 10.1038/ni.1645

    Figure Lengend Snippet: Expression of rapamycin-sensitive mTOR pathway components in pDCs. ( a ) Immunoblot analysis of phosphorylated (phospho-) and total mTOR in lysates of RAW cells (4 × 10 6 ) transiently transfected for 40 h with cyan fluorescent protein–tagged MyD88 and IRF7-YFP, treated with vehicle (−) or rapamycin (+) and then stimulated for 0–60 min with CpG-A–DOTAP. Results are representative of two independent experiments. ( b ) Immunoblot analysis of phosphorylated and total mTOR, p70S6K, 4E-BP1 and Akt in lysates of purified pDCs (1 × 10 6 ) stimulated for 15 min with CpG-A in the presence (Rap) or absence (Medium) of rapamycin. Results are representative of three independent experiments.

    Article Snippet: CsA, FK506 (tacrolimus) and rapamycin (sirolimus) were from Sigma.

    Techniques: Expressing, Transfection, Purification

    Rapamycin does not affect TLR9 expression or interaction with CpG-A. ( a ) Flow cytometry of isolated spleen pDCs stimulated with CpG-A with or without rapamycin pretreatment, then stained for intracellular TLR9. ( b ) Deconvolution microscopy of RAW cells expressing YFP-tagged TLR9 (green), pretreated with vehicle (left) or rapamycin (right) and then incubated for 90 min with indodicarbocyanine-tagged CpG-A–DOTAP (red; 5 μg/ml). Two images were obtained for each condition. Data are representative of two independent experiments.

    Journal: Nature immunology

    Article Title: Toll-like receptor-mediated induction of type I interferon in plasmacytoid dendritic cells requires the rapamycin-sensitive PI(3)K-mTOR-p70S6K pathway

    doi: 10.1038/ni.1645

    Figure Lengend Snippet: Rapamycin does not affect TLR9 expression or interaction with CpG-A. ( a ) Flow cytometry of isolated spleen pDCs stimulated with CpG-A with or without rapamycin pretreatment, then stained for intracellular TLR9. ( b ) Deconvolution microscopy of RAW cells expressing YFP-tagged TLR9 (green), pretreated with vehicle (left) or rapamycin (right) and then incubated for 90 min with indodicarbocyanine-tagged CpG-A–DOTAP (red; 5 μg/ml). Two images were obtained for each condition. Data are representative of two independent experiments.

    Article Snippet: CsA, FK506 (tacrolimus) and rapamycin (sirolimus) were from Sigma.

    Techniques: Expressing, Flow Cytometry, Cytometry, Isolation, Staining, Microscopy, Incubation

    In vivo administration of rapamycin inhibits YF-17D-induced production of IFN-α in the serum. ( a ) ELISA of IFN-α production by pDCs from C57BL/6 mice treated daily for 3 d with soluble rapamycin (1.5 mg per kg body weight per day) or rapamycin encapsulated in PLGA microparticles (2 mg per mouse per day), then vaccinated subcutaneously with YF-17D on day 4; blood and spleens were obtained at various times after injection for analysis of serum (right) and pDCs enriched from spleens with microbeads coated with anti–mouse PDCA1 (left). * P

    Journal: Nature immunology

    Article Title: Toll-like receptor-mediated induction of type I interferon in plasmacytoid dendritic cells requires the rapamycin-sensitive PI(3)K-mTOR-p70S6K pathway

    doi: 10.1038/ni.1645

    Figure Lengend Snippet: In vivo administration of rapamycin inhibits YF-17D-induced production of IFN-α in the serum. ( a ) ELISA of IFN-α production by pDCs from C57BL/6 mice treated daily for 3 d with soluble rapamycin (1.5 mg per kg body weight per day) or rapamycin encapsulated in PLGA microparticles (2 mg per mouse per day), then vaccinated subcutaneously with YF-17D on day 4; blood and spleens were obtained at various times after injection for analysis of serum (right) and pDCs enriched from spleens with microbeads coated with anti–mouse PDCA1 (left). * P

    Article Snippet: CsA, FK506 (tacrolimus) and rapamycin (sirolimus) were from Sigma.

    Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Mouse Assay, Injection

    Rapamycin inhibits TLR-mediated IFN-α secretion by pDCs. ( a ) ELISA of IFN-α in supernatants of isolated pDCs pretreated for 3 h with various doses of rapamycin (Rap) and then stimulated with CpG-A (10 μg/ml; left) or treated with vehicle (Veh) or rapamycin and then stimulated with various doses of CpG-A (right). ( b ) ELISA of IFN-β, TNF, IL-6 and CXCL10 in supernatants of purified mouse pDCs pretreated with various doses of rapamycin and then stimulated for 24 h with CpG-A (10 μg/ml). ( c ) ELISA of IFN-α in supernatants of purified spleen pDCs cultured for 24 h with CpG-A (10 μg/ml), loxoribine (Loxo; 1 mM), YF-17D or vesicular stomatitis virus (VSV; all viruses at a multiplicity of infection of 1) after pretreatment with rapamycin (Rap) or vehicle. ( d ) Apoptotic death of isolated mouse pDCs pretreated with vehicle or 20 nM or 500 nM rapamycin, assessed by staining with annexin V and propidium iodide. Numbers in quadrants indicate percent cells in each. ( e ) Proliferation of and cytokine induction in naive CD4 + CD62L + OT-II T cells (1 × 10 5 cells/ well) cultured with pDCs (5 × 10 4 ) pretreated with ovalbumin peptide (10 μg/ml) and CpG-A (10 μg/ml) plus vehicle or rapamycin (100 nM for 3 h), assessed after 3 d by incorporation of [ 3 H]thymidine (top left; proliferation) and ELISA of IFN-γ, IL-4, IL-10, IL-13 and IL-17. *, P

    Journal: Nature immunology

    Article Title: Toll-like receptor-mediated induction of type I interferon in plasmacytoid dendritic cells requires the rapamycin-sensitive PI(3)K-mTOR-p70S6K pathway

    doi: 10.1038/ni.1645

    Figure Lengend Snippet: Rapamycin inhibits TLR-mediated IFN-α secretion by pDCs. ( a ) ELISA of IFN-α in supernatants of isolated pDCs pretreated for 3 h with various doses of rapamycin (Rap) and then stimulated with CpG-A (10 μg/ml; left) or treated with vehicle (Veh) or rapamycin and then stimulated with various doses of CpG-A (right). ( b ) ELISA of IFN-β, TNF, IL-6 and CXCL10 in supernatants of purified mouse pDCs pretreated with various doses of rapamycin and then stimulated for 24 h with CpG-A (10 μg/ml). ( c ) ELISA of IFN-α in supernatants of purified spleen pDCs cultured for 24 h with CpG-A (10 μg/ml), loxoribine (Loxo; 1 mM), YF-17D or vesicular stomatitis virus (VSV; all viruses at a multiplicity of infection of 1) after pretreatment with rapamycin (Rap) or vehicle. ( d ) Apoptotic death of isolated mouse pDCs pretreated with vehicle or 20 nM or 500 nM rapamycin, assessed by staining with annexin V and propidium iodide. Numbers in quadrants indicate percent cells in each. ( e ) Proliferation of and cytokine induction in naive CD4 + CD62L + OT-II T cells (1 × 10 5 cells/ well) cultured with pDCs (5 × 10 4 ) pretreated with ovalbumin peptide (10 μg/ml) and CpG-A (10 μg/ml) plus vehicle or rapamycin (100 nM for 3 h), assessed after 3 d by incorporation of [ 3 H]thymidine (top left; proliferation) and ELISA of IFN-γ, IL-4, IL-10, IL-13 and IL-17. *, P

    Article Snippet: CsA, FK506 (tacrolimus) and rapamycin (sirolimus) were from Sigma.

    Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Purification, Cell Culture, Infection, Staining

    Blood concentrations and biliary excretion of (A,C) sirolimus and (B,D) tacrolimus after daily treatment over 2 weeks with sirolimus (SRL, 1 mg kg −1 p.o.), tacrolimus (TRL 1 mg kg −1 i.p.) and of SRL (1 mg kg −1 p.o.) plus CyA (10 mg kg −1 i.p.) or TRL (1 mg kg −1 i.p.). Means±s.e.mean of 10 animals each. * Significant difference ( P

    Journal: British Journal of Pharmacology

    Article Title: Sirolimus/cyclosporine/tacrolimus interactions on bile flow and biliary excretion of immunosuppressants in a subchronic bile fistula rat model

    doi: 10.1038/sj.bjp.0704756

    Figure Lengend Snippet: Blood concentrations and biliary excretion of (A,C) sirolimus and (B,D) tacrolimus after daily treatment over 2 weeks with sirolimus (SRL, 1 mg kg −1 p.o.), tacrolimus (TRL 1 mg kg −1 i.p.) and of SRL (1 mg kg −1 p.o.) plus CyA (10 mg kg −1 i.p.) or TRL (1 mg kg −1 i.p.). Means±s.e.mean of 10 animals each. * Significant difference ( P

    Article Snippet: Sirolimus (Rapamune®) was obtained from Wyeth-Pharma GmbH, Münster, Germany, as a solution (1 mg ml−1 ) for oral application.

    Techniques:

    Blood concentrations of (A) cyclosporine and (B) its metabolites after daily treatment over 2 weeks with cyclosporine (CyA, 10 mg kg −1 i.p.) or sirolimus (SRL, 1 mg kg −1 p.o.) plus CyA (10 mg kg −1 i.p.). Means±s.e.mean of 10 animals each. * Significant difference ( P

    Journal: British Journal of Pharmacology

    Article Title: Sirolimus/cyclosporine/tacrolimus interactions on bile flow and biliary excretion of immunosuppressants in a subchronic bile fistula rat model

    doi: 10.1038/sj.bjp.0704756

    Figure Lengend Snippet: Blood concentrations of (A) cyclosporine and (B) its metabolites after daily treatment over 2 weeks with cyclosporine (CyA, 10 mg kg −1 i.p.) or sirolimus (SRL, 1 mg kg −1 p.o.) plus CyA (10 mg kg −1 i.p.). Means±s.e.mean of 10 animals each. * Significant difference ( P

    Article Snippet: Sirolimus (Rapamune®) was obtained from Wyeth-Pharma GmbH, Münster, Germany, as a solution (1 mg ml−1 ) for oral application.

    Techniques:

    Bile flow and biliary excretion of bile salts and cholesterol after daily treatment over 2 weeks with sirolimus (SRL, 1 mg kg −1 p.o.), cyclosporine (CyA, 10 mg kg −1 i.p.), tacrolimus (TRL, 1 mg kg −1 i.p.) and of SRL (1 mg kg −1 p.o.) plus CyA (10 mg kg −1 i.p.) or TRL (1 mg kg −1 i.p.). Means±s.e.mean of 10 animals each. * Significant difference ( P

    Journal: British Journal of Pharmacology

    Article Title: Sirolimus/cyclosporine/tacrolimus interactions on bile flow and biliary excretion of immunosuppressants in a subchronic bile fistula rat model

    doi: 10.1038/sj.bjp.0704756

    Figure Lengend Snippet: Bile flow and biliary excretion of bile salts and cholesterol after daily treatment over 2 weeks with sirolimus (SRL, 1 mg kg −1 p.o.), cyclosporine (CyA, 10 mg kg −1 i.p.), tacrolimus (TRL, 1 mg kg −1 i.p.) and of SRL (1 mg kg −1 p.o.) plus CyA (10 mg kg −1 i.p.) or TRL (1 mg kg −1 i.p.). Means±s.e.mean of 10 animals each. * Significant difference ( P

    Article Snippet: Sirolimus (Rapamune®) was obtained from Wyeth-Pharma GmbH, Münster, Germany, as a solution (1 mg ml−1 ) for oral application.

    Techniques: Flow Cytometry

    TS-1/22 plus basiliximab and sirolimus prolongs islet allograft survival.

    Journal: The Journal of Clinical Investigation

    Article Title: LFA-1-specific therapy prolongs allograft survival in rhesus macaques

    doi: 10.1172/JCI43895

    Figure Lengend Snippet: TS-1/22 plus basiliximab and sirolimus prolongs islet allograft survival.

    Article Snippet: Basiliximab (Simulect, Novartis) was purchased from the Emory University Hospital pharmacy, and sirolimus (Rapamune) was purchased from LC Laboratories.

    Techniques:

    MoSNT2 is associated with the MoTor signaling pathway. (A) Vegetative growth of M. oryzae on CM agar medium supplemented with or without 1 μg/ml rapamycin (rapa.). (B) Inhibition rate of rapamycin on the mycelial growth. (C) Expression profiles of MoSNT2 and MoTOR in the wild-type Guy11 strain at different developmental processes. (D) Linear correlation between qRT-PCR-measured expression levels of MoSNT2 and MoTOR . (E) qRT-PCR analysis of MoSNT2 expression levels in the Guy11 strain in response to rapamycin. The Guy11 strain grown in liquid CM for 48 h was transferred into fresh liquid CM in the presence or absence of 1 μg/ml rapamycin for 6 h before total RNA extraction.

    Journal: Autophagy

    Article Title: MoSnt2-dependent deacetylation of histone H3 mediates MoTor-dependent autophagy and plant infection by the rice blast fungus Magnaporthe oryzae

    doi: 10.1080/15548627.2018.1458171

    Figure Lengend Snippet: MoSNT2 is associated with the MoTor signaling pathway. (A) Vegetative growth of M. oryzae on CM agar medium supplemented with or without 1 μg/ml rapamycin (rapa.). (B) Inhibition rate of rapamycin on the mycelial growth. (C) Expression profiles of MoSNT2 and MoTOR in the wild-type Guy11 strain at different developmental processes. (D) Linear correlation between qRT-PCR-measured expression levels of MoSNT2 and MoTOR . (E) qRT-PCR analysis of MoSNT2 expression levels in the Guy11 strain in response to rapamycin. The Guy11 strain grown in liquid CM for 48 h was transferred into fresh liquid CM in the presence or absence of 1 μg/ml rapamycin for 6 h before total RNA extraction.

    Article Snippet: A rapamycin (Selleckchem, S1039) stock solution was prepared in DMSO (Amresco, 0231) at a concentration of 10 mg/ml.

    Techniques: Inhibition, Expressing, Quantitative RT-PCR, RNA Extraction

    Effects of rapamycin on vegetative growth, autophagic cell death and pathogenicity of M. oryzae . (A) Plate colonies of the Guy11 strain in CM agar medium. The Guy11 strain was grown on CM medium supplemented with rapamycin (rapa.) at the indicated concentration. Solvent DMSO was seperately added into medium as a control. (B) Diameters of plate colonies recorded every 2 days. (C) Autophagic conidial cell death of the Guy11 strain at 24 hpi of appressorium development on hydrophobic coverslip in the presence of rapamycin. Scale bar: 10 μm. (D) Percentage of Guy11 strain spores containing 3 totally-collapsed conidial cells at 24 hpi (n > 100, triple replications, ** P

    Journal: Autophagy

    Article Title: MoSnt2-dependent deacetylation of histone H3 mediates MoTor-dependent autophagy and plant infection by the rice blast fungus Magnaporthe oryzae

    doi: 10.1080/15548627.2018.1458171

    Figure Lengend Snippet: Effects of rapamycin on vegetative growth, autophagic cell death and pathogenicity of M. oryzae . (A) Plate colonies of the Guy11 strain in CM agar medium. The Guy11 strain was grown on CM medium supplemented with rapamycin (rapa.) at the indicated concentration. Solvent DMSO was seperately added into medium as a control. (B) Diameters of plate colonies recorded every 2 days. (C) Autophagic conidial cell death of the Guy11 strain at 24 hpi of appressorium development on hydrophobic coverslip in the presence of rapamycin. Scale bar: 10 μm. (D) Percentage of Guy11 strain spores containing 3 totally-collapsed conidial cells at 24 hpi (n > 100, triple replications, ** P

    Article Snippet: A rapamycin (Selleckchem, S1039) stock solution was prepared in DMSO (Amresco, 0231) at a concentration of 10 mg/ml.

    Techniques: Concentration Assay

    Effect of mTOR inhibitors on cell growth. G-415 ( A ) and TGBC-2TKB ( B ) cells were treated with LY294002, rapamycin, RAD001, and AZD8055 at shown concentrations. Notes: Cell viability was measured after 24, 48, and 72 hours of treatment using CellTiter 96® AQueous One Solution Reagent assay. Viable cells are expressed as the percentage of control (untreated cells; 0.01% dimethylsulfoxide). Values represent the mean ± standard error of the mean of at least three independent experiments in triplicate. * P

    Journal: OncoTargets and therapy

    Article Title: AKT/mTOR substrate P70S6K is frequently phosphorylated in gallbladder cancer tissue and cell lines

    doi: 10.2147/OTT.S46897

    Figure Lengend Snippet: Effect of mTOR inhibitors on cell growth. G-415 ( A ) and TGBC-2TKB ( B ) cells were treated with LY294002, rapamycin, RAD001, and AZD8055 at shown concentrations. Notes: Cell viability was measured after 24, 48, and 72 hours of treatment using CellTiter 96® AQueous One Solution Reagent assay. Viable cells are expressed as the percentage of control (untreated cells; 0.01% dimethylsulfoxide). Values represent the mean ± standard error of the mean of at least three independent experiments in triplicate. * P

    Article Snippet: Rapamycin (sirolimus, #S1039), everolimus (RAD001, #S1120), and AZD8055 (#S1555) were purchased from Selleck Chemicals (Houston, TX, USA).

    Techniques:

    Effect of mTOR inhibitors of mTOR on AKT/mTOR signaling pathway in two gallbladder cancer cell lines. G-415 and TGBC-2TKB cells were grown in Roswell Park Memorial Institute 1640 medium supplemented with 10% fetal bovine serum and treated with LY294002 (10 μM), rapamycin (50 nM), RAD001 (1 nM), and AZD5085 (25 nM) for 18 hours. Control cells received an equivalent amount of solvent (0.1% dimethylsulfoxide). Western blot analysis was carried out using antibodies against the total and phosphorylated portion of AKT, p70S6K, 4E-BP1, and eIF4E proteins. Abbreviations: DMSO, dimethyl sulfoxide; LY29, LY294002 PI3 kinase inhibitor; Rapa, rapamycin; Everol, everolimus; AZD, AZD-8055.

    Journal: OncoTargets and therapy

    Article Title: AKT/mTOR substrate P70S6K is frequently phosphorylated in gallbladder cancer tissue and cell lines

    doi: 10.2147/OTT.S46897

    Figure Lengend Snippet: Effect of mTOR inhibitors of mTOR on AKT/mTOR signaling pathway in two gallbladder cancer cell lines. G-415 and TGBC-2TKB cells were grown in Roswell Park Memorial Institute 1640 medium supplemented with 10% fetal bovine serum and treated with LY294002 (10 μM), rapamycin (50 nM), RAD001 (1 nM), and AZD5085 (25 nM) for 18 hours. Control cells received an equivalent amount of solvent (0.1% dimethylsulfoxide). Western blot analysis was carried out using antibodies against the total and phosphorylated portion of AKT, p70S6K, 4E-BP1, and eIF4E proteins. Abbreviations: DMSO, dimethyl sulfoxide; LY29, LY294002 PI3 kinase inhibitor; Rapa, rapamycin; Everol, everolimus; AZD, AZD-8055.

    Article Snippet: Rapamycin (sirolimus, #S1039), everolimus (RAD001, #S1120), and AZD8055 (#S1555) were purchased from Selleck Chemicals (Houston, TX, USA).

    Techniques: Western Blot

    Effect of rapamycin, RAD001, and AZD8055 on cell migration. Notes: Assays were performed using 24-well Transwell™ plates containing polycarbonate filters with an 8 μm pore size. Before seeding, G-415 and TGBC-2TKB cells were exposed to LY294002 (10 μM), rapamycin (50 nM), RAD001 (1 nM), and AZD8055 (25 nM) for 1 hour. Dimethylsulfoxide 0.1% was used as the control. Cells were counted after 24 hours using an optic microscope in 16 randomly selected fields. Results were obtained from three separate experiments and are expressed as the mean ± standard error of the mean. *** P

    Journal: OncoTargets and therapy

    Article Title: AKT/mTOR substrate P70S6K is frequently phosphorylated in gallbladder cancer tissue and cell lines

    doi: 10.2147/OTT.S46897

    Figure Lengend Snippet: Effect of rapamycin, RAD001, and AZD8055 on cell migration. Notes: Assays were performed using 24-well Transwell™ plates containing polycarbonate filters with an 8 μm pore size. Before seeding, G-415 and TGBC-2TKB cells were exposed to LY294002 (10 μM), rapamycin (50 nM), RAD001 (1 nM), and AZD8055 (25 nM) for 1 hour. Dimethylsulfoxide 0.1% was used as the control. Cells were counted after 24 hours using an optic microscope in 16 randomly selected fields. Results were obtained from three separate experiments and are expressed as the mean ± standard error of the mean. *** P

    Article Snippet: Rapamycin (sirolimus, #S1039), everolimus (RAD001, #S1120), and AZD8055 (#S1555) were purchased from Selleck Chemicals (Houston, TX, USA).

    Techniques: Migration, Microscopy

    Activation of AKT/mTOR pathway in human gallbladder cancer cell lines. Total protein (35 μg) from eight gallbladder cancer cell lines were subjected to immunoblotting using antibodies directed against AKT, phospho-AKT, p70S6K, phospho-p70S6K, eIF4E, phospho-eIF4E, 4E-BP1, and phospho-4E-BP1. Protein loading was normalized using an antibody recognizing β-actin. Abbreviations: kDa, kiloDalton; β-actin, beta actin; AKT, protein kinase B; mTOR, mammalian target of rapamycin; p70S6K, p70S6 kinase; eIF4E, eukaryotic translation initiation factor 4E; 4E-BP1, eukaryotic translation initiation factor 4e-binding protein 1.

    Journal: OncoTargets and therapy

    Article Title: AKT/mTOR substrate P70S6K is frequently phosphorylated in gallbladder cancer tissue and cell lines

    doi: 10.2147/OTT.S46897

    Figure Lengend Snippet: Activation of AKT/mTOR pathway in human gallbladder cancer cell lines. Total protein (35 μg) from eight gallbladder cancer cell lines were subjected to immunoblotting using antibodies directed against AKT, phospho-AKT, p70S6K, phospho-p70S6K, eIF4E, phospho-eIF4E, 4E-BP1, and phospho-4E-BP1. Protein loading was normalized using an antibody recognizing β-actin. Abbreviations: kDa, kiloDalton; β-actin, beta actin; AKT, protein kinase B; mTOR, mammalian target of rapamycin; p70S6K, p70S6 kinase; eIF4E, eukaryotic translation initiation factor 4E; 4E-BP1, eukaryotic translation initiation factor 4e-binding protein 1.

    Article Snippet: Rapamycin (sirolimus, #S1039), everolimus (RAD001, #S1120), and AZD8055 (#S1555) were purchased from Selleck Chemicals (Houston, TX, USA).

    Techniques: Activation Assay, Binding Assay

    Rab escort protein1 (REP1) depletion suppresses cell growth and survival. ( A ) MiaPaCa2 cells were transfected with control (CTL) and REP1 small interfering RNAs (siRNAs). After 48 h, immunoblotting was performed to analyze REP1 protein levels. Cells were treated with CTL and REP1 siRNAs and incubated for 24 h. Then, the cells were transfected with Flag-REP1 plasmid additionally and further incubated in the IncuCyte TM for monitoring cell proliferation. At the 72-h incubation time point, cell confluence levels were presented as a percentage using the IncuCyte TM analyzer. ( B , C ) MiaPaCa2 cells were transfected with control and REP1siRNAs, which replaced the following day with serum-, glucose-, or glutamine-free medium and then incubated for another 24 h.Cell morphology was observed by brightfield image. Scale bar: 50 μm ( B ). Cell death was assessed by using the Annexin V/propidium iodide (PI) assay ( C ). Error bars indicate mean +/− standard error for n = 3 independent experiments. ( D ) MiaPaCa2 cells were transfected with control (CTL) or REP1siRNAs, which replaced the following day with 1 µM rapamycin and then further incubated for monitoring cell confluence using IncuCyte TM . At 72 h time point, cell confluence levels were presented as percentage. Statistical significance was determined via t-test; * p

    Journal: International Journal of Molecular Sciences

    Article Title: REP1 Modulates Autophagy and Macropinocytosis to Enhance Cancer Cell Survival

    doi: 10.3390/ijms18091866

    Figure Lengend Snippet: Rab escort protein1 (REP1) depletion suppresses cell growth and survival. ( A ) MiaPaCa2 cells were transfected with control (CTL) and REP1 small interfering RNAs (siRNAs). After 48 h, immunoblotting was performed to analyze REP1 protein levels. Cells were treated with CTL and REP1 siRNAs and incubated for 24 h. Then, the cells were transfected with Flag-REP1 plasmid additionally and further incubated in the IncuCyte TM for monitoring cell proliferation. At the 72-h incubation time point, cell confluence levels were presented as a percentage using the IncuCyte TM analyzer. ( B , C ) MiaPaCa2 cells were transfected with control and REP1siRNAs, which replaced the following day with serum-, glucose-, or glutamine-free medium and then incubated for another 24 h.Cell morphology was observed by brightfield image. Scale bar: 50 μm ( B ). Cell death was assessed by using the Annexin V/propidium iodide (PI) assay ( C ). Error bars indicate mean +/− standard error for n = 3 independent experiments. ( D ) MiaPaCa2 cells were transfected with control (CTL) or REP1siRNAs, which replaced the following day with 1 µM rapamycin and then further incubated for monitoring cell confluence using IncuCyte TM . At 72 h time point, cell confluence levels were presented as percentage. Statistical significance was determined via t-test; * p

    Article Snippet: The protease inhibitor cocktail tablet was from Roche Applied Bioscience, Penzberg, Upper Bavaria, Germany Rapamycin (S1039) and Torin2 were purchased from Selleck Chemicals (Houston, TX, USA).

    Techniques: Transfection, CTL Assay, Incubation, Plasmid Preparation

    Effects of rapamycin (Rap) and 3-methyladenine (3-MA) on carotid blood partial pressure of oxygen (PO 2 ) (a), Lung injury score (b), and WET/DRY ratio (c) in control and diabetic rats undergoing myocardial ischemia reperfusion (IR). All values are expressed as means ± S.D., n = 8. ∗ P

    Journal: Journal of Diabetes Research

    Article Title: The Roles of Autophagy in Acute Lung Injury Induced by Myocardial Ischemia Reperfusion in Diabetic Rats

    doi: 10.1155/2018/5047526

    Figure Lengend Snippet: Effects of rapamycin (Rap) and 3-methyladenine (3-MA) on carotid blood partial pressure of oxygen (PO 2 ) (a), Lung injury score (b), and WET/DRY ratio (c) in control and diabetic rats undergoing myocardial ischemia reperfusion (IR). All values are expressed as means ± S.D., n = 8. ∗ P

    Article Snippet: Diabetic rats were either pretreated with 3-methyladenine (3-MA, 15 mg/kg, intraperitoneally) [ ] 30 min before ischemia or intravenous rapamycin (Rap, 0.25 mg/kg) [ ] (Selleck Chemicals, USA) 15 min before ischemia or equal volume of saline for control.

    Techniques:

    Effects of rapamycin (Rap) and 3-methyladenine (3-MA) on the pulmonary expression of LC-3II and Beclin-1 in diabetic rats. Control (C) and diabetic rats (D) pretreated with intraperitoneal 3-MA (15 mg/kg) 30 min or intravenous Rap (0.25 mg/kg) 15 min before ischemia or equal volume of saline were subjected to 30 min of myocardial ischemia followed by 2 h of reperfusion. (a) Representative Western blot demonstrating LC-3II, LC-3I, and Beclin-1 expression with GAPDH as loading control. (b) LC-3II/LC-3I ratio was calculated by relative densitometric values and expressed as fold to C group. (c) Beclin-1 express was calculated by relative densitometric values and expressed as fold to C group. All values are expressed as mean means ± S.D., n = 8. ∗ P

    Journal: Journal of Diabetes Research

    Article Title: The Roles of Autophagy in Acute Lung Injury Induced by Myocardial Ischemia Reperfusion in Diabetic Rats

    doi: 10.1155/2018/5047526

    Figure Lengend Snippet: Effects of rapamycin (Rap) and 3-methyladenine (3-MA) on the pulmonary expression of LC-3II and Beclin-1 in diabetic rats. Control (C) and diabetic rats (D) pretreated with intraperitoneal 3-MA (15 mg/kg) 30 min or intravenous Rap (0.25 mg/kg) 15 min before ischemia or equal volume of saline were subjected to 30 min of myocardial ischemia followed by 2 h of reperfusion. (a) Representative Western blot demonstrating LC-3II, LC-3I, and Beclin-1 expression with GAPDH as loading control. (b) LC-3II/LC-3I ratio was calculated by relative densitometric values and expressed as fold to C group. (c) Beclin-1 express was calculated by relative densitometric values and expressed as fold to C group. All values are expressed as mean means ± S.D., n = 8. ∗ P

    Article Snippet: Diabetic rats were either pretreated with 3-methyladenine (3-MA, 15 mg/kg, intraperitoneally) [ ] 30 min before ischemia or intravenous rapamycin (Rap, 0.25 mg/kg) [ ] (Selleck Chemicals, USA) 15 min before ischemia or equal volume of saline for control.

    Techniques: Expressing, Western Blot

    Effects of rapamycin (Rap) and 3-methyladenine (3-MA) on leukocyte count (a), TNF- α (b), IL-6 (c); IL-8 (d) in BAL fluid; and SOD (e) and 15-F2t-Isop (f) in lung tissues from control and diabetic rats undergoing myocardial ischemia reperfusion (IR). All values are expressed as means ± S.D., n = 8. ∗ P

    Journal: Journal of Diabetes Research

    Article Title: The Roles of Autophagy in Acute Lung Injury Induced by Myocardial Ischemia Reperfusion in Diabetic Rats

    doi: 10.1155/2018/5047526

    Figure Lengend Snippet: Effects of rapamycin (Rap) and 3-methyladenine (3-MA) on leukocyte count (a), TNF- α (b), IL-6 (c); IL-8 (d) in BAL fluid; and SOD (e) and 15-F2t-Isop (f) in lung tissues from control and diabetic rats undergoing myocardial ischemia reperfusion (IR). All values are expressed as means ± S.D., n = 8. ∗ P

    Article Snippet: Diabetic rats were either pretreated with 3-methyladenine (3-MA, 15 mg/kg, intraperitoneally) [ ] 30 min before ischemia or intravenous rapamycin (Rap, 0.25 mg/kg) [ ] (Selleck Chemicals, USA) 15 min before ischemia or equal volume of saline for control.

    Techniques:

    Measurement of the autophagic flux in ZIKV-infected HUVEC. ( A ) Western blot analysis of p62 degradation. The time course of expression of p62 in ZIKV-infected HUVEC at MOI 1 was investigated using anti-p62 antibody. Mock-infected HUVEC were used as negative controls, rapamycin (100 nM) treatment was used as a positive control, and GAPDH was used as a protein-loading control; ( B ) the ratios of P62 to GAPDH were calculated, and representative results are presented with graphs. Data are presented as means from three independent experiments. Compared to the control group, significance is analyzed with two-tailed Student’s t test. * p

    Journal: Viruses

    Article Title: Zika Virus Induces Autophagy in Human Umbilical Vein Endothelial Cells

    doi: 10.3390/v10050259

    Figure Lengend Snippet: Measurement of the autophagic flux in ZIKV-infected HUVEC. ( A ) Western blot analysis of p62 degradation. The time course of expression of p62 in ZIKV-infected HUVEC at MOI 1 was investigated using anti-p62 antibody. Mock-infected HUVEC were used as negative controls, rapamycin (100 nM) treatment was used as a positive control, and GAPDH was used as a protein-loading control; ( B ) the ratios of P62 to GAPDH were calculated, and representative results are presented with graphs. Data are presented as means from three independent experiments. Compared to the control group, significance is analyzed with two-tailed Student’s t test. * p

    Article Snippet: Antibodies and Chemicals Rapamycin and wortmannin were purchased from Selleck (Shanghai, China).

    Techniques: Infection, Western Blot, Expressing, Positive Control, Two Tailed Test

    Treatment of rapamycin has no effect on ZIKV infection in HUVEC. ( A ) HUVEC were pretreated with rapamycin (Rapa) (100 nM) or DMSO (Ctrl) in complete medium and then processed as described for Figure 4 A; ( B ) At 18 and 24 hpi, extracellular virus yields were determined by plaque assay on Vero cells and expressed as pfu/mL; ( C ) HUVEC were pretreated with Rapa or DMSO and then processed as described for panel A, with drug concentrations of 50, 100, 200, and 500 nM; ( D ) Extracellular virus yields at 24 hpi were determined and expressed as pfu/mL. Data are presented as means ± SDs from three independent experiments. Significance was analyzed with two-tailed Student’s t test and compared to the control group. # p > 0.05.

    Journal: Viruses

    Article Title: Zika Virus Induces Autophagy in Human Umbilical Vein Endothelial Cells

    doi: 10.3390/v10050259

    Figure Lengend Snippet: Treatment of rapamycin has no effect on ZIKV infection in HUVEC. ( A ) HUVEC were pretreated with rapamycin (Rapa) (100 nM) or DMSO (Ctrl) in complete medium and then processed as described for Figure 4 A; ( B ) At 18 and 24 hpi, extracellular virus yields were determined by plaque assay on Vero cells and expressed as pfu/mL; ( C ) HUVEC were pretreated with Rapa or DMSO and then processed as described for panel A, with drug concentrations of 50, 100, 200, and 500 nM; ( D ) Extracellular virus yields at 24 hpi were determined and expressed as pfu/mL. Data are presented as means ± SDs from three independent experiments. Significance was analyzed with two-tailed Student’s t test and compared to the control group. # p > 0.05.

    Article Snippet: Antibodies and Chemicals Rapamycin and wortmannin were purchased from Selleck (Shanghai, China).

    Techniques: Infection, Plaque Assay, Two Tailed Test

    ZIKV induces autophagy in HUVEC. ( A ) Confocal microscopy. HUVEC were infected with lentivirus expressing mTagRFP-mWasabi-LC3 followed by treatment at 48 h post-infection with mock treatment as a negative control, rapamycin (100 nM) treatment as a positive control, or infection of ZIKV (MOI = 1) for 24 h. The cells were observed by confocal microscopy with scale bars indicating 10 µm; ( B ) percent of cells with LC3 dots from total LC3-expressing cells was calculated. Data were derived from at least 100 cells for each sample. * p

    Journal: Viruses

    Article Title: Zika Virus Induces Autophagy in Human Umbilical Vein Endothelial Cells

    doi: 10.3390/v10050259

    Figure Lengend Snippet: ZIKV induces autophagy in HUVEC. ( A ) Confocal microscopy. HUVEC were infected with lentivirus expressing mTagRFP-mWasabi-LC3 followed by treatment at 48 h post-infection with mock treatment as a negative control, rapamycin (100 nM) treatment as a positive control, or infection of ZIKV (MOI = 1) for 24 h. The cells were observed by confocal microscopy with scale bars indicating 10 µm; ( B ) percent of cells with LC3 dots from total LC3-expressing cells was calculated. Data were derived from at least 100 cells for each sample. * p

    Article Snippet: Antibodies and Chemicals Rapamycin and wortmannin were purchased from Selleck (Shanghai, China).

    Techniques: Confocal Microscopy, Infection, Expressing, Negative Control, Positive Control, Derivative Assay

    ( a ) Western blots of targeted clones show significant differences in collapsin response mediator protein B2 (CRMP2B) levels (64 kDa) between the two isogenic cell groups. ( b ) In the presence of 30 nM Rapamycin, CRMPZB is reduced for both cell groups, but most dramatically in the 13PNR cells. ( c ) Quantification of western blots using the ImageJ software. * P

    Journal: Translational Psychiatry

    Article Title: The DPYSL2 gene connects mTOR and schizophrenia

    doi: 10.1038/tp.2016.204

    Figure Lengend Snippet: ( a ) Western blots of targeted clones show significant differences in collapsin response mediator protein B2 (CRMP2B) levels (64 kDa) between the two isogenic cell groups. ( b ) In the presence of 30 nM Rapamycin, CRMPZB is reduced for both cell groups, but most dramatically in the 13PNR cells. ( c ) Quantification of western blots using the ImageJ software. * P

    Article Snippet: For Rapamycin exposures, we grew cells in standard media containing 30nM Rapamycin (SelleckChem, Houston, TX, USA, cat. # S1039) for 24 hours.

    Techniques: Western Blot, Clone Assay, Software

    Addition of IL-15 recovers rapamycin-mediated abnormal DETC homeostasis. ( a,b ) 6–8 weeks wild-type male C57BL/6J mice were administered rapamycin or vehicle control daily for 8 weeks. At 8 weeks after administration of the vehicle control and at, 2, 4, and 8 weeks after rapamycin treatment, epidermal sheets were obtained for assessing the number of DETCs by FACS (panel a) and the production of IL-15 by Western analysis (panel b). ( c ) IL-15 (1 μg) or vehicle control was intradermally injected daily on the back skin of rapamycin-treated mice for 3 days, and epidermal sheets were excised for assessing the number of DETCs by FACS. All data represent at least three independent experiments and each experiment includes 6 animals (3 control and 3 experimental mice). All error bars represent mean ± SEM. * P

    Journal: Scientific Reports

    Article Title: Weakened IL-15 Production and Impaired mTOR Activation Alter Dendritic Epidermal T Cell Homeostasis in Diabetic Mice

    doi: 10.1038/s41598-017-05950-5

    Figure Lengend Snippet: Addition of IL-15 recovers rapamycin-mediated abnormal DETC homeostasis. ( a,b ) 6–8 weeks wild-type male C57BL/6J mice were administered rapamycin or vehicle control daily for 8 weeks. At 8 weeks after administration of the vehicle control and at, 2, 4, and 8 weeks after rapamycin treatment, epidermal sheets were obtained for assessing the number of DETCs by FACS (panel a) and the production of IL-15 by Western analysis (panel b). ( c ) IL-15 (1 μg) or vehicle control was intradermally injected daily on the back skin of rapamycin-treated mice for 3 days, and epidermal sheets were excised for assessing the number of DETCs by FACS. All data represent at least three independent experiments and each experiment includes 6 animals (3 control and 3 experimental mice). All error bars represent mean ± SEM. * P

    Article Snippet: For rapamycin administration, mice were injected i.p. daily with 200 μl of vehicle control or 1% rapamycin (Selleck Chemicals, Houston) in 0.2% carboxymethyl cellulose and 0.25% Tween 80 (Sigma-Aldrich, USA) in distilled H2 O.

    Techniques: Mouse Assay, FACS, Western Blot, Injection

    IGF-1 regulates keratinocyte-derived IL-15 in an mTOR-dependent manner. ( a,b ) Primary keratinocytes were isolated from newborn C57BL/6J male wild-type mice, and CD11c + cells were depleted by MACS. These cells were cultured for 1 day in 0, 20, 40, 80, or 100 ng/ml IGF-1. IL-15 was assessed at the mRNA level by real-time PCR (panel a) and at the protein level by Western blotting (panel b). ( c ) Phosphorylated and total S6K, and Akt in keratinocytes were measured in the presence or absence of IGF-1 (100 ng/ml, for 24 hours) by Western blotting. ( d ) Analysis of IGF-1-induced IL-15 production suppression by rapamycin in keratinocytes after 24 h culture. All data represent at least three independent experiments. All error bars represent mean ± SEM. * P

    Journal: Scientific Reports

    Article Title: Weakened IL-15 Production and Impaired mTOR Activation Alter Dendritic Epidermal T Cell Homeostasis in Diabetic Mice

    doi: 10.1038/s41598-017-05950-5

    Figure Lengend Snippet: IGF-1 regulates keratinocyte-derived IL-15 in an mTOR-dependent manner. ( a,b ) Primary keratinocytes were isolated from newborn C57BL/6J male wild-type mice, and CD11c + cells were depleted by MACS. These cells were cultured for 1 day in 0, 20, 40, 80, or 100 ng/ml IGF-1. IL-15 was assessed at the mRNA level by real-time PCR (panel a) and at the protein level by Western blotting (panel b). ( c ) Phosphorylated and total S6K, and Akt in keratinocytes were measured in the presence or absence of IGF-1 (100 ng/ml, for 24 hours) by Western blotting. ( d ) Analysis of IGF-1-induced IL-15 production suppression by rapamycin in keratinocytes after 24 h culture. All data represent at least three independent experiments. All error bars represent mean ± SEM. * P

    Article Snippet: For rapamycin administration, mice were injected i.p. daily with 200 μl of vehicle control or 1% rapamycin (Selleck Chemicals, Houston) in 0.2% carboxymethyl cellulose and 0.25% Tween 80 (Sigma-Aldrich, USA) in distilled H2 O.

    Techniques: Derivative Assay, Isolation, Mouse Assay, Magnetic Cell Separation, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot

    Both productions of IGF-1 and IL-15 in epidermis are regulated by mTOR pathway. ( a ) 6–8 weeks wild-type male C57BL/6 J mice were administered STZ daily for 6 days. At day 0 (no treatment), 3, 6, 9, and 12 after STZ treatment, epidermal sheets were obtained for detecting the production of IGF-1 by Western blotting. ( b ) Wild-type C57BL/6J mice were administered rapamycin daily for 8 weeks. At 0, 2, 4, and 8 weeks after rapamycin treatment, epidermal sheets were obtained for detecting the production of IGF-1 by Western blotting. ( c ) The kinetics of IL-15 and IGF-1 in rapamycin-treated mice is shown. ( d ) The kinetics of IL-15 and IGF-1 in STZ-treated mice is shown. All data represent at least three independent experiments and each experiment includes 6 animals (3 control and 3 experimental mice). All error bars represent mean ± SEM. * P

    Journal: Scientific Reports

    Article Title: Weakened IL-15 Production and Impaired mTOR Activation Alter Dendritic Epidermal T Cell Homeostasis in Diabetic Mice

    doi: 10.1038/s41598-017-05950-5

    Figure Lengend Snippet: Both productions of IGF-1 and IL-15 in epidermis are regulated by mTOR pathway. ( a ) 6–8 weeks wild-type male C57BL/6 J mice were administered STZ daily for 6 days. At day 0 (no treatment), 3, 6, 9, and 12 after STZ treatment, epidermal sheets were obtained for detecting the production of IGF-1 by Western blotting. ( b ) Wild-type C57BL/6J mice were administered rapamycin daily for 8 weeks. At 0, 2, 4, and 8 weeks after rapamycin treatment, epidermal sheets were obtained for detecting the production of IGF-1 by Western blotting. ( c ) The kinetics of IL-15 and IGF-1 in rapamycin-treated mice is shown. ( d ) The kinetics of IL-15 and IGF-1 in STZ-treated mice is shown. All data represent at least three independent experiments and each experiment includes 6 animals (3 control and 3 experimental mice). All error bars represent mean ± SEM. * P

    Article Snippet: For rapamycin administration, mice were injected i.p. daily with 200 μl of vehicle control or 1% rapamycin (Selleck Chemicals, Houston) in 0.2% carboxymethyl cellulose and 0.25% Tween 80 (Sigma-Aldrich, USA) in distilled H2 O.

    Techniques: Mouse Assay, Western Blot

    Rapamycin treatment reduced seizures and number of mitochondria. A. Frequency and duration of seizures (sz) in cKO mice in the absence or presence of rapamycin, n = 5 in each group. Data in A are expressed as mean ± SD. Unpaired t -test versus vehicle-treated mice. B. Histoimmunostaining analyses of mTOR signaling (p-S6), mitochondria (COXIV), and neuronal excitation (c-FOS). Scale bars: 100 μm. WT, wild-type. C. The fluorescence intensity was quantified by ImageJ. n = 5 in each group. Data in C are expressed as mean ± SD. One-way ANOVA test, followed by Dunnett's post hoc test for multiple comparisons (WT mice versus cKO mice; cKO mice versus Rapamycin-treated cKO mice (cKO_Rapa); WT mice versus cKO_Rapa mice) was performed and adjusted P values were calculated. * p

    Journal: PLoS ONE

    Article Title: Epilepsy in a melanocyte-lineage mTOR hyperactivation mouse model: A novel epilepsy model

    doi: 10.1371/journal.pone.0228204

    Figure Lengend Snippet: Rapamycin treatment reduced seizures and number of mitochondria. A. Frequency and duration of seizures (sz) in cKO mice in the absence or presence of rapamycin, n = 5 in each group. Data in A are expressed as mean ± SD. Unpaired t -test versus vehicle-treated mice. B. Histoimmunostaining analyses of mTOR signaling (p-S6), mitochondria (COXIV), and neuronal excitation (c-FOS). Scale bars: 100 μm. WT, wild-type. C. The fluorescence intensity was quantified by ImageJ. n = 5 in each group. Data in C are expressed as mean ± SD. One-way ANOVA test, followed by Dunnett's post hoc test for multiple comparisons (WT mice versus cKO mice; cKO mice versus Rapamycin-treated cKO mice (cKO_Rapa); WT mice versus cKO_Rapa mice) was performed and adjusted P values were calculated. * p

    Article Snippet: In the rapamycin treatment experiments, behavior analysis was performed after 3 weeks of oral sirolimus (Selleck) in distilled water.

    Techniques: Mouse Assay, Fluorescence

    The effects of combined treatment on the growth of tumor in vivo . Mice with SKOV3 endometrial xenografts (200 mm 3 ) were randomized into treatment with vehicle, BGJ398 (10 or 30 mg/kg), Rapamycin (0.4 or 1.0 mg/kg), or a combination of both inhibitors. A. Tumor growth in low-dose BGJ398 group (with or without Rapamycin). B. The growth of tumor in high-dose BGJ398 group (with or without Rapamycin). C. The expression levels of p-S6, p-Akt, p-PI3K and p-ERK1/2 in mice treated with 30 mg/kg BGJ398 (with or without 1.0 mg/kg rapamycin). D. The network of FGFR signaling pathway. Inhibition of FGFR results in the phosphorylation of FRS2-α and activation of downstream molecules such as ERK1/2 and PI3K, and the PI3K/AKT pathway activates the mTOR/S6 signalling, which is negatively regulated by rapamycin.

    Journal: American Journal of Translational Research

    Article Title: Combined inhibition of FGFR and mTOR pathways is effective in suppressing ovarian cancer

    doi:

    Figure Lengend Snippet: The effects of combined treatment on the growth of tumor in vivo . Mice with SKOV3 endometrial xenografts (200 mm 3 ) were randomized into treatment with vehicle, BGJ398 (10 or 30 mg/kg), Rapamycin (0.4 or 1.0 mg/kg), or a combination of both inhibitors. A. Tumor growth in low-dose BGJ398 group (with or without Rapamycin). B. The growth of tumor in high-dose BGJ398 group (with or without Rapamycin). C. The expression levels of p-S6, p-Akt, p-PI3K and p-ERK1/2 in mice treated with 30 mg/kg BGJ398 (with or without 1.0 mg/kg rapamycin). D. The network of FGFR signaling pathway. Inhibition of FGFR results in the phosphorylation of FRS2-α and activation of downstream molecules such as ERK1/2 and PI3K, and the PI3K/AKT pathway activates the mTOR/S6 signalling, which is negatively regulated by rapamycin.

    Article Snippet: Inhibitors of FGFR (BGJ398) and mTOR (Rapamycin), paraformaldehyde and dimethyl sulphoxide (DMSO) were purchased from Selleck Chemicals (Houston, Texas, USA).

    Techniques: In Vivo, Mouse Assay, Expressing, Inhibition, Activation Assay

    Combination of rapamycin and EF24 induces mitochondrial dysfunction by ROS/JNK signaling pathways. ( A ) Treatment of cells with combination of 5 µM rapamycin and 2 µM EF24 decreased mitochondrial membrane potential (Δψm). Cells were treated for 12 h and then stained with JC-1. Mitochondrial dysfunction was not evident when cells were pretreated with NAC or catalase [JC-1 accumulation in the mitochondria appear red, non-accumulating JC-1 as a monomer appear green; scale bar=20 μm]. ( B ) Effect of NAC on SGC-7901 mitochondrial morphology following combined rapamycin and EF24 treatment. ( C ) EF24 enhanced rapamycin-induced p-JNK levels. NAC or catalase pretreatment reversed the effects of 12-h combined rapamycin and EF24 in SGC-7901 and BGC-823 cells. The densitometric quantification bar graphs are shown in Supplementary file. Data were collected from three independent experiments and representative images were shown.

    Journal: Redox Biology

    Article Title: Synergistic antitumor activity of rapamycin and EF24 via increasing ROS for the treatment of gastric cancer

    doi: 10.1016/j.redox.2016.09.006

    Figure Lengend Snippet: Combination of rapamycin and EF24 induces mitochondrial dysfunction by ROS/JNK signaling pathways. ( A ) Treatment of cells with combination of 5 µM rapamycin and 2 µM EF24 decreased mitochondrial membrane potential (Δψm). Cells were treated for 12 h and then stained with JC-1. Mitochondrial dysfunction was not evident when cells were pretreated with NAC or catalase [JC-1 accumulation in the mitochondria appear red, non-accumulating JC-1 as a monomer appear green; scale bar=20 μm]. ( B ) Effect of NAC on SGC-7901 mitochondrial morphology following combined rapamycin and EF24 treatment. ( C ) EF24 enhanced rapamycin-induced p-JNK levels. NAC or catalase pretreatment reversed the effects of 12-h combined rapamycin and EF24 in SGC-7901 and BGC-823 cells. The densitometric quantification bar graphs are shown in Supplementary file. Data were collected from three independent experiments and representative images were shown.

    Article Snippet: 2.1 Cell culture and reagents Rapamycin was purchased from Selleck Chemicals (Houston, TX).

    Techniques: Staining

    EF24 enhances the anti-tumor activity of rapamycin in human gastric cancer xenografts. SGC-7901 cells were injected in nude mice. Mice were then treated with rapamycin, EF24, or a combination of both. Figure showing tumor weight ( A ) and tumor volume ( B and C ) [*p

    Journal: Redox Biology

    Article Title: Synergistic antitumor activity of rapamycin and EF24 via increasing ROS for the treatment of gastric cancer

    doi: 10.1016/j.redox.2016.09.006

    Figure Lengend Snippet: EF24 enhances the anti-tumor activity of rapamycin in human gastric cancer xenografts. SGC-7901 cells were injected in nude mice. Mice were then treated with rapamycin, EF24, or a combination of both. Figure showing tumor weight ( A ) and tumor volume ( B and C ) [*p

    Article Snippet: 2.1 Cell culture and reagents Rapamycin was purchased from Selleck Chemicals (Houston, TX).

    Techniques: Activity Assay, Injection, Mouse Assay

    EF24 enhances rapamycin-induced ER stress response by modulating ROS levels . ( A ) Western blot analysis of endoplasmic reticulum (ER) stress related proteins in cells following treatment with rapamycin and EF24 [p-eIF2α=phosphorylated eukaryotic initiation factor 2α; ATF4=activating transcription factor-4; p-PERK=phosphorylated protein kinase RNA-like endoplasmic reticulum kinase; and CHOP=CCAAT/enhancer-binding protein homologous protein]. ( B ) NAC and catalase reversed the combined treatment-induced ER stress response in cells as assessed by western blot analysis. ( C ) Effect of combined rapamycin and EF24 treatment on the morphology of endoplasmic reticulum in SGC-7901 cells. SGC-7901 cells were pre-treated 5 mM NAC for 2 h before exposure to 5 µM rapamycin in combination with 2 µM EF24 for 6 h [×10,000 or ×20,000]. The densitometric quantification bar graphs are shown in Supplementary file. Data were collected from three independent experiments and representative images were shown.

    Journal: Redox Biology

    Article Title: Synergistic antitumor activity of rapamycin and EF24 via increasing ROS for the treatment of gastric cancer

    doi: 10.1016/j.redox.2016.09.006

    Figure Lengend Snippet: EF24 enhances rapamycin-induced ER stress response by modulating ROS levels . ( A ) Western blot analysis of endoplasmic reticulum (ER) stress related proteins in cells following treatment with rapamycin and EF24 [p-eIF2α=phosphorylated eukaryotic initiation factor 2α; ATF4=activating transcription factor-4; p-PERK=phosphorylated protein kinase RNA-like endoplasmic reticulum kinase; and CHOP=CCAAT/enhancer-binding protein homologous protein]. ( B ) NAC and catalase reversed the combined treatment-induced ER stress response in cells as assessed by western blot analysis. ( C ) Effect of combined rapamycin and EF24 treatment on the morphology of endoplasmic reticulum in SGC-7901 cells. SGC-7901 cells were pre-treated 5 mM NAC for 2 h before exposure to 5 µM rapamycin in combination with 2 µM EF24 for 6 h [×10,000 or ×20,000]. The densitometric quantification bar graphs are shown in Supplementary file. Data were collected from three independent experiments and representative images were shown.

    Article Snippet: 2.1 Cell culture and reagents Rapamycin was purchased from Selleck Chemicals (Houston, TX).

    Techniques: Western Blot, Binding Assay

    Rapamycin causes cytotoxicity inhuman gastric cancer cells and normal cells. ( A ) Cytotoxic effect of rapamycin in human gastric cancer cells and normal GES-1 cells as assessed by cell viability. SGC-7901, BGC-823 and GES-1 cells were incubated with increasing doses of rapamycin (0.31–100 μM) for 24 h. Cell viability was determined by MTT assay. ( B ) Induction of apoptosis in human gastric cancer cells was determined by Annexin V/PI flow cytometry following treatment with rapamycin (10, 20 or 40 μM) for 24 h. ( C ) Quantification of apoptotic cells presented as percent total [*p

    Journal: Redox Biology

    Article Title: Synergistic antitumor activity of rapamycin and EF24 via increasing ROS for the treatment of gastric cancer

    doi: 10.1016/j.redox.2016.09.006

    Figure Lengend Snippet: Rapamycin causes cytotoxicity inhuman gastric cancer cells and normal cells. ( A ) Cytotoxic effect of rapamycin in human gastric cancer cells and normal GES-1 cells as assessed by cell viability. SGC-7901, BGC-823 and GES-1 cells were incubated with increasing doses of rapamycin (0.31–100 μM) for 24 h. Cell viability was determined by MTT assay. ( B ) Induction of apoptosis in human gastric cancer cells was determined by Annexin V/PI flow cytometry following treatment with rapamycin (10, 20 or 40 μM) for 24 h. ( C ) Quantification of apoptotic cells presented as percent total [*p

    Article Snippet: 2.1 Cell culture and reagents Rapamycin was purchased from Selleck Chemicals (Houston, TX).

    Techniques: Incubation, MTT Assay, Flow Cytometry, Cytometry

    EF24 enhances rapamycin-induced ROS generation. ( A and B ) Rapamycin-induced ROS generation was measured in SGC-7901 cells and BGC-823 cells by staining with DCFH-DA. ( A ) SGC-7901 and BGC-823 cells were treated with 10 μM rapamycin for different time periods. ( B ) Cells treated with different concentrations of rapamycin for 1 h. ( C and D ) EF24 enhances rapamycin-induced ROS generation in SGC-7901 ( C ) and BGC-823 ( D ) cells. Cells were treated with EF24, rapamycin, or a combination of both for 1 h. Intracellular ROS generation was inhibited with pretreatment of 5 mM NAC for 2 h. Right bar graphs show quantification of flow cytometry data [*p

    Journal: Redox Biology

    Article Title: Synergistic antitumor activity of rapamycin and EF24 via increasing ROS for the treatment of gastric cancer

    doi: 10.1016/j.redox.2016.09.006

    Figure Lengend Snippet: EF24 enhances rapamycin-induced ROS generation. ( A and B ) Rapamycin-induced ROS generation was measured in SGC-7901 cells and BGC-823 cells by staining with DCFH-DA. ( A ) SGC-7901 and BGC-823 cells were treated with 10 μM rapamycin for different time periods. ( B ) Cells treated with different concentrations of rapamycin for 1 h. ( C and D ) EF24 enhances rapamycin-induced ROS generation in SGC-7901 ( C ) and BGC-823 ( D ) cells. Cells were treated with EF24, rapamycin, or a combination of both for 1 h. Intracellular ROS generation was inhibited with pretreatment of 5 mM NAC for 2 h. Right bar graphs show quantification of flow cytometry data [*p

    Article Snippet: 2.1 Cell culture and reagents Rapamycin was purchased from Selleck Chemicals (Houston, TX).

    Techniques: Staining, Flow Cytometry, Cytometry

    EF24 enhances the anti-tumor activity of rapamycin in gastric cancer cell lines. ( A-B ) SGC-7901,BGC-823 and GES-1 cells were pretreated with 0.5 μM EF24 and then exposed to increasing doses of rapamycin (0.31–100 μM) for 24 h. Cell viability was determined by MTT assay. ( C ) EF24 enhances rapamycin-induced apoptosis in SGC-7901 and BGC-823 cells as assessed by AnnexinV/PI staining. ( D ) Quantification of apoptotic cells [*p

    Journal: Redox Biology

    Article Title: Synergistic antitumor activity of rapamycin and EF24 via increasing ROS for the treatment of gastric cancer

    doi: 10.1016/j.redox.2016.09.006

    Figure Lengend Snippet: EF24 enhances the anti-tumor activity of rapamycin in gastric cancer cell lines. ( A-B ) SGC-7901,BGC-823 and GES-1 cells were pretreated with 0.5 μM EF24 and then exposed to increasing doses of rapamycin (0.31–100 μM) for 24 h. Cell viability was determined by MTT assay. ( C ) EF24 enhances rapamycin-induced apoptosis in SGC-7901 and BGC-823 cells as assessed by AnnexinV/PI staining. ( D ) Quantification of apoptotic cells [*p

    Article Snippet: 2.1 Cell culture and reagents Rapamycin was purchased from Selleck Chemicals (Houston, TX).

    Techniques: Activity Assay, MTT Assay, Staining

    Synergistic effect rapamycin and EF24 is dependent on ROS generation. ( A ) NAC or catalase pretreatment normalized the synergistic effect of rapamycin and EF24 in SGC-7901 and BGC-823 cells. Cells were pretreated with 5 mM NAC or 2000 U/mL catalase for 2 h before exposure to the combination treatment of rapamycin and EF24 for 24 h. Apoptotic cells were determined by AnnexinV/PI staining. ( B ) Quantification of Annexin V/PI flow cytometry data [*p

    Journal: Redox Biology

    Article Title: Synergistic antitumor activity of rapamycin and EF24 via increasing ROS for the treatment of gastric cancer

    doi: 10.1016/j.redox.2016.09.006

    Figure Lengend Snippet: Synergistic effect rapamycin and EF24 is dependent on ROS generation. ( A ) NAC or catalase pretreatment normalized the synergistic effect of rapamycin and EF24 in SGC-7901 and BGC-823 cells. Cells were pretreated with 5 mM NAC or 2000 U/mL catalase for 2 h before exposure to the combination treatment of rapamycin and EF24 for 24 h. Apoptotic cells were determined by AnnexinV/PI staining. ( B ) Quantification of Annexin V/PI flow cytometry data [*p

    Article Snippet: 2.1 Cell culture and reagents Rapamycin was purchased from Selleck Chemicals (Houston, TX).

    Techniques: Staining, Flow Cytometry, Cytometry

    Rapamycin (Rapa) attenuates beige fat gene expression in primary adipocytes. A and B : Expression of thermogenic genes, Ucp1 and Elovl3 , in primary adipocytes treated with 1 μmol/L CL ( A ) or 1 mmol/L 8-Br-cAMP (cAMP) ( B ) for 24 h with or without 500 nmol/L rapamycin pretreatment for 24 h. C and D : Adrb3 gene expression in the same cells treated with CL ( C ) or 8-Br-cAMP ( D ) with or without rapamycin pretreatment. E : PKA substrate phosphorylation (pPKA), phosphorylated CREB (pCREB), and pS6 in fully differentiated adipocytes treated with rapamycin for 24 h or PKA inhibitor for 1 h before treatment with CL for 20 min. Values shown are mean ± SEM. n = 4–6. * P

    Journal: Diabetes

    Article Title: Rapamycin Blocks Induction of the Thermogenic Program in White Adipose Tissue

    doi: 10.2337/db15-0502

    Figure Lengend Snippet: Rapamycin (Rapa) attenuates beige fat gene expression in primary adipocytes. A and B : Expression of thermogenic genes, Ucp1 and Elovl3 , in primary adipocytes treated with 1 μmol/L CL ( A ) or 1 mmol/L 8-Br-cAMP (cAMP) ( B ) for 24 h with or without 500 nmol/L rapamycin pretreatment for 24 h. C and D : Adrb3 gene expression in the same cells treated with CL ( C ) or 8-Br-cAMP ( D ) with or without rapamycin pretreatment. E : PKA substrate phosphorylation (pPKA), phosphorylated CREB (pCREB), and pS6 in fully differentiated adipocytes treated with rapamycin for 24 h or PKA inhibitor for 1 h before treatment with CL for 20 min. Values shown are mean ± SEM. n = 4–6. * P

    Article Snippet: Fully differentiated adipocytes were treated with 500 nmol/L rapamycin (Selleckchem) for 24 h or H-89 (Cayman Chemical) for 1 h before addition of either 1 μmol/L CL or 1 mmol/L cAMP analog (8-Br-cAMP) (Sigma) for 24 h.

    Techniques: Expressing