5g4 mab 6c5  (HyTest)


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    Structured Review

    HyTest 5g4 mab 6c5
    5g4 Mab 6c5, supplied by HyTest, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5g4 mab 6c5/product/HyTest
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    5g4 mab 6c5 - by Bioz Stars, 2019-06
    86/100 stars

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    Related Articles

    Homogenization:

    Article Title: Axial Tubule Junctions Activate Atrial Ca2+ Release Across Species
    Article Snippet: Then, atrial tissues were homogenized in ice-cold homogenization buffer [HEPES 10 mM, sucrose 300 mM, NaCl 150 mM, EGTA 1 mM, CaCl2 2 mM, Triton X-100 0.5% (v/v), protease and phosphatase inhibitor mix (Roche), pH 7.4] using a Miccra D-1 homogenizer. .. Membranes were blocked for 1 h in 5% (w/v) non-fat milk in Tris-buffered saline with 0.05% (v/v) Tween 20, and incubated with the primary antibodies overnight at 4°C as follows: RyR2 1:2500 (HPA020028, Sigma-Aldrich); NCX 1:1000 (11-13, Swant); SERCA2 1:2000 (A010-20, Badrilla); Na,K-ATPase α1 subunit 1:500 (sc-21712, Santa-Cruz); GAPDH 1:160,000 (5G4 Mab 6C5, HyTest); PLN 1:2500 (ab2865, Abcam); PLM 1:1000 (13721-1-AP, Proteintech).

    Incubation:

    Article Title: Axial Tubule Junctions Activate Atrial Ca2+ Release Across Species
    Article Snippet: Proteins were transferred onto PVDF membranes (0.45 μm, Immobilon-FL, Merck Millipore) using the Bio-Rad criterion blotter (plate electrodes). .. Membranes were blocked for 1 h in 5% (w/v) non-fat milk in Tris-buffered saline with 0.05% (v/v) Tween 20, and incubated with the primary antibodies overnight at 4°C as follows: RyR2 1:2500 (HPA020028, Sigma-Aldrich); NCX 1:1000 (11-13, Swant); SERCA2 1:2000 (A010-20, Badrilla); Na,K-ATPase α1 subunit 1:500 (sc-21712, Santa-Cruz); GAPDH 1:160,000 (5G4 Mab 6C5, HyTest); PLN 1:2500 (ab2865, Abcam); PLM 1:1000 (13721-1-AP, Proteintech). .. After washing, blots were incubated with fluorescence-labeled anti-rabbit or anti-mouse secondary antibodies at a dilution of 1:10,000 for a minimum period of 1 h at room temperature (LI-COR, P/N 926-32212, P/N 926-68072, P/N 926-32213, P/N 926-68073).

    other:

    Article Title: Axial Tubule Junctions Activate Atrial Ca2+ Release Across Species
    Article Snippet: Membranes were blocked for 1 h in 5% (w/v) non-fat milk in Tris-buffered saline with 0.05% (v/v) Tween 20, and incubated with the primary antibodies overnight at 4°C as follows: RyR2 1:2500 (HPA020028, Sigma-Aldrich); NCX 1:1000 (11-13, Swant); SERCA2 1:2000 (A010-20, Badrilla); Na,K-ATPase α1 subunit 1:500 (sc-21712, Santa-Cruz); GAPDH 1:160,000 (5G4 Mab 6C5, HyTest); PLN 1:2500 (ab2865, Abcam); PLM 1:1000 (13721-1-AP, Proteintech).

    Imaging:

    Article Title: Axial Tubule Junctions Activate Atrial Ca2+ Release Across Species
    Article Snippet: Membranes were blocked for 1 h in 5% (w/v) non-fat milk in Tris-buffered saline with 0.05% (v/v) Tween 20, and incubated with the primary antibodies overnight at 4°C as follows: RyR2 1:2500 (HPA020028, Sigma-Aldrich); NCX 1:1000 (11-13, Swant); SERCA2 1:2000 (A010-20, Badrilla); Na,K-ATPase α1 subunit 1:500 (sc-21712, Santa-Cruz); GAPDH 1:160,000 (5G4 Mab 6C5, HyTest); PLN 1:2500 (ab2865, Abcam); PLM 1:1000 (13721-1-AP, Proteintech). .. After washing, blots were incubated with fluorescence-labeled anti-rabbit or anti-mouse secondary antibodies at a dilution of 1:10,000 for a minimum period of 1 h at room temperature (LI-COR, P/N 926-32212, P/N 926-68072, P/N 926-32213, P/N 926-68073).

    BIA-KA:

    Article Title: Axial Tubule Junctions Activate Atrial Ca2+ Release Across Species
    Article Snippet: Protein concentrations were determined with the Pierce BCA protein Assay Kit (Thermo Fisher Scientific). .. Membranes were blocked for 1 h in 5% (w/v) non-fat milk in Tris-buffered saline with 0.05% (v/v) Tween 20, and incubated with the primary antibodies overnight at 4°C as follows: RyR2 1:2500 (HPA020028, Sigma-Aldrich); NCX 1:1000 (11-13, Swant); SERCA2 1:2000 (A010-20, Badrilla); Na,K-ATPase α1 subunit 1:500 (sc-21712, Santa-Cruz); GAPDH 1:160,000 (5G4 Mab 6C5, HyTest); PLN 1:2500 (ab2865, Abcam); PLM 1:1000 (13721-1-AP, Proteintech).

    Western Blot:

    Article Title: Axial Tubule Junctions Activate Atrial Ca2+ Release Across Species
    Article Snippet: For Western blots 20 μg of protein per lane were resolved by SDS-PAGE using 4–20% Tris-HCl protein gels (3450033, Bio-Rad). .. Membranes were blocked for 1 h in 5% (w/v) non-fat milk in Tris-buffered saline with 0.05% (v/v) Tween 20, and incubated with the primary antibodies overnight at 4°C as follows: RyR2 1:2500 (HPA020028, Sigma-Aldrich); NCX 1:1000 (11-13, Swant); SERCA2 1:2000 (A010-20, Badrilla); Na,K-ATPase α1 subunit 1:500 (sc-21712, Santa-Cruz); GAPDH 1:160,000 (5G4 Mab 6C5, HyTest); PLN 1:2500 (ab2865, Abcam); PLM 1:1000 (13721-1-AP, Proteintech).

    SDS Page:

    Article Title: Axial Tubule Junctions Activate Atrial Ca2+ Release Across Species
    Article Snippet: For Western blots 20 μg of protein per lane were resolved by SDS-PAGE using 4–20% Tris-HCl protein gels (3450033, Bio-Rad). .. Membranes were blocked for 1 h in 5% (w/v) non-fat milk in Tris-buffered saline with 0.05% (v/v) Tween 20, and incubated with the primary antibodies overnight at 4°C as follows: RyR2 1:2500 (HPA020028, Sigma-Aldrich); NCX 1:1000 (11-13, Swant); SERCA2 1:2000 (A010-20, Badrilla); Na,K-ATPase α1 subunit 1:500 (sc-21712, Santa-Cruz); GAPDH 1:160,000 (5G4 Mab 6C5, HyTest); PLN 1:2500 (ab2865, Abcam); PLM 1:1000 (13721-1-AP, Proteintech).

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  • 97
    HyTest gapdh
    Molecular determinants of ryanodine receptor type-2 (RyR2) dysfunction. (A) Representative Western blot examples of total RyR2, Ser2808-, and Ser2814-phosphorylated RyR2 and calsequestrin (CSQ; left), as well as <t>junctin</t> and junctophilin-2 (JPH2; right) protein expression levels. <t>GAPDH</t> was used as loading control. (B) Quantification of total RyR2, Ser2808-, and Ser2814-phosphorylated RyR2, Ser2808/total RyR2, and Ser2814/total RyR2 protein expression levels in Ctl (white bars), HFrEF (blue bars) and HFrEF-cAF (red/blue-striped bars) patients. (C) Protein expression levels of the RyR2-interacting proteins CSQ, junctin, and JPH2 in Ctl, HFrEF and HFrEF-cAF patients. Data are shown relative to Ctl. Numbers in bars indicate number of patients. ∗ indicates P
    Gapdh, supplied by HyTest, used in various techniques. Bioz Stars score: 97/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gapdh/product/HyTest
    Average 97 stars, based on 20 article reviews
    Price from $9.99 to $1999.99
    gapdh - by Bioz Stars, 2019-06
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    79
    HyTest housekeeping gene glyceraldehyde 3 phosphate dehydrogenase gapdh
    Everolimus (EV) inhibits cancer cell growth in vitro and in vivo. a The murine melanoma cell line B16-F10 and the human breast cancer cell lines MCF-7 and MDA-MB-231 were treated with EV in a dose-dependent (0, 1, 10, and 100 nM) and time-dependent (0, 24, 48, and 72 h) manner. Cell viability was assessed with the CellTiter-Blue® assay. b Western blots used to assess the ability of EV concentrations to inhibit the phosphorylation of the mammalian target of rapamycin (mTOR) protein and p70 S6 kinase after 24 h of treatment in the cell lines investigated. Glyceraldehyde 3-phosphate dehydrogenase ( <t>GAPDH</t> ) is shown as the housekeeping control. c Female immunocompetent (C57BL/6) and immunocompromised (NMRI nude) mice were inoculated subcutaneously with B16-F10 and MDA-MB-231 cells, respectively. Tumor growth was assessed after daily treatment with 1 mg/kg of EV for 2 and 4 weeks in each respective model. In vitro and in vivo data are shown as mean ± SD of at least three independent experiments or ten mice per group, respectively. Cell viability assays were analyzed for each time point using two-way analysis of variance and in vivo data by Student’s t test. Significance between EV treatments and the control condition was apparent only at 72 h and is indicated by asterisks on the graphs. In the B16-F10 graph, EV treatment was significant at inhibiting viability only at 72 h at concentrations of 10 and 100 nM. In the MDA-MB-231 and MCF-7 graphs, all concentrations of EV were significant to the same degree. ** p
    Housekeeping Gene Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by HyTest, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/housekeeping gene glyceraldehyde 3 phosphate dehydrogenase gapdh/product/HyTest
    Average 79 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    Molecular determinants of ryanodine receptor type-2 (RyR2) dysfunction. (A) Representative Western blot examples of total RyR2, Ser2808-, and Ser2814-phosphorylated RyR2 and calsequestrin (CSQ; left), as well as junctin and junctophilin-2 (JPH2; right) protein expression levels. GAPDH was used as loading control. (B) Quantification of total RyR2, Ser2808-, and Ser2814-phosphorylated RyR2, Ser2808/total RyR2, and Ser2814/total RyR2 protein expression levels in Ctl (white bars), HFrEF (blue bars) and HFrEF-cAF (red/blue-striped bars) patients. (C) Protein expression levels of the RyR2-interacting proteins CSQ, junctin, and JPH2 in Ctl, HFrEF and HFrEF-cAF patients. Data are shown relative to Ctl. Numbers in bars indicate number of patients. ∗ indicates P

    Journal: Frontiers in Physiology

    Article Title: Profibrotic, Electrical, and Calcium-Handling Remodeling of the Atria in Heart Failure Patients With and Without Atrial Fibrillation

    doi: 10.3389/fphys.2018.01383

    Figure Lengend Snippet: Molecular determinants of ryanodine receptor type-2 (RyR2) dysfunction. (A) Representative Western blot examples of total RyR2, Ser2808-, and Ser2814-phosphorylated RyR2 and calsequestrin (CSQ; left), as well as junctin and junctophilin-2 (JPH2; right) protein expression levels. GAPDH was used as loading control. (B) Quantification of total RyR2, Ser2808-, and Ser2814-phosphorylated RyR2, Ser2808/total RyR2, and Ser2814/total RyR2 protein expression levels in Ctl (white bars), HFrEF (blue bars) and HFrEF-cAF (red/blue-striped bars) patients. (C) Protein expression levels of the RyR2-interacting proteins CSQ, junctin, and JPH2 in Ctl, HFrEF and HFrEF-cAF patients. Data are shown relative to Ctl. Numbers in bars indicate number of patients. ∗ indicates P

    Article Snippet: Protein levels of α smooth-muscle actin (αSMA, 1:500, A5228, Sigma-Aldrich, St. Louis, MO, United States), calsequestrin (CSQ, 1:2,500, PA1-913, Thermo Fisher Scientific), collagen 1α (Col1a, 1:1,000, sc-293182, Santa Cruz Biotechnology, Santa Cruz, CA, United States), connexin-40 (Cx40, 1:1,000, ab1726, Merck Millipore, Burlington, MA, United States), total and Ser368-phosphorylated connexin-43 (Cx43, 1:1,000, 3511, Cell Signaling Technology, Danvers, MA, United States), fibronectin (1:1,000, sc-8422, Santa Cruz Biotechnology), GAPDH (1:20,000, 5G4 6C5, HyTest, Turku, Finland), junctin (1:2,000, LS-C196703, LifeSpan BioSciences, Seattle, WA, United States), junctophilin-2 (1:1,000, sc-134875, Santa Cruz), matrix metallopeptidase 9 (MMP9, 1:200, ab38898, Abcam, Cambridge, United Kingdom), Na+ -Ca2+ -exchanger type-1 (NCX1, 1:1,000, R3F1, Swant, Marly, Switzerland), periostin (1:1,000, sc-134875, Santa Cruz Biotechnology), total, Ser16- and Thr17-phosphorylated phospholamban (PLB, all 1:1,000, ab2865 and ab92697, Abcam, Cambridge, United Kingdom and A010-13, Badrilla Ltd., Leeds, United Kingdom), total ryanodine receptor type-2 channel (RyR2, 1:1,000, MA3-916, Thermo Fisher Scientific), sarcolipin (1:100, ABT13, Merck Millipore), sarcoplasmic reticulum (SR) Ca2+ -ATPase type-2a (SERCA2a; 1:2,000, sc-8095, Santa Cruz Biotechnology), transforming growth factor β1 (TGF-β1, 1:1000, ab9758, Abcam), and vimentin (1:1,000, sc373717, Santa Cruz Biotechnology) were determined using appropriate primary antibodies.

    Techniques: Western Blot, Expressing, CTL Assay

    Atrial profibrotic and connexin remodeling. (A) Representative Western blot examples (top) and quantification of protein expression (bottom; mean ± SEM) of periostin, vimentin, α-smooth muscle actin (α-SMA), matrix metallopeptidase 9 (MMP9), transforming growth factor-β1 (TGF-β1), fibronectin and collagen 1α (col1a) in right-atrial tissue homogenates from Ctl (white bars), HFrEF (blue bars) or HFrEF-cAF (red/blue-striped bars) patients. Vertical white lines separate non-adjacent lanes on the same blot. (B) Representative Western blot examples (top) and quantification of protein expression (bottom; mean ± SEM) of total connexin-40 (Cx40), total and Ser368-phosphorylated connexin-43 (Cx43). GAPDH was used as loading control and is shown for the samples used for Western blots of periostin, vimentin, α-SMA and MMP9, for TGF-β1, and for fibronectin and col1a. Numbers in bars indicate number of patients. ∗ indicates P

    Journal: Frontiers in Physiology

    Article Title: Profibrotic, Electrical, and Calcium-Handling Remodeling of the Atria in Heart Failure Patients With and Without Atrial Fibrillation

    doi: 10.3389/fphys.2018.01383

    Figure Lengend Snippet: Atrial profibrotic and connexin remodeling. (A) Representative Western blot examples (top) and quantification of protein expression (bottom; mean ± SEM) of periostin, vimentin, α-smooth muscle actin (α-SMA), matrix metallopeptidase 9 (MMP9), transforming growth factor-β1 (TGF-β1), fibronectin and collagen 1α (col1a) in right-atrial tissue homogenates from Ctl (white bars), HFrEF (blue bars) or HFrEF-cAF (red/blue-striped bars) patients. Vertical white lines separate non-adjacent lanes on the same blot. (B) Representative Western blot examples (top) and quantification of protein expression (bottom; mean ± SEM) of total connexin-40 (Cx40), total and Ser368-phosphorylated connexin-43 (Cx43). GAPDH was used as loading control and is shown for the samples used for Western blots of periostin, vimentin, α-SMA and MMP9, for TGF-β1, and for fibronectin and col1a. Numbers in bars indicate number of patients. ∗ indicates P

    Article Snippet: Protein levels of α smooth-muscle actin (αSMA, 1:500, A5228, Sigma-Aldrich, St. Louis, MO, United States), calsequestrin (CSQ, 1:2,500, PA1-913, Thermo Fisher Scientific), collagen 1α (Col1a, 1:1,000, sc-293182, Santa Cruz Biotechnology, Santa Cruz, CA, United States), connexin-40 (Cx40, 1:1,000, ab1726, Merck Millipore, Burlington, MA, United States), total and Ser368-phosphorylated connexin-43 (Cx43, 1:1,000, 3511, Cell Signaling Technology, Danvers, MA, United States), fibronectin (1:1,000, sc-8422, Santa Cruz Biotechnology), GAPDH (1:20,000, 5G4 6C5, HyTest, Turku, Finland), junctin (1:2,000, LS-C196703, LifeSpan BioSciences, Seattle, WA, United States), junctophilin-2 (1:1,000, sc-134875, Santa Cruz), matrix metallopeptidase 9 (MMP9, 1:200, ab38898, Abcam, Cambridge, United Kingdom), Na+ -Ca2+ -exchanger type-1 (NCX1, 1:1,000, R3F1, Swant, Marly, Switzerland), periostin (1:1,000, sc-134875, Santa Cruz Biotechnology), total, Ser16- and Thr17-phosphorylated phospholamban (PLB, all 1:1,000, ab2865 and ab92697, Abcam, Cambridge, United Kingdom and A010-13, Badrilla Ltd., Leeds, United Kingdom), total ryanodine receptor type-2 channel (RyR2, 1:1,000, MA3-916, Thermo Fisher Scientific), sarcolipin (1:100, ABT13, Merck Millipore), sarcoplasmic reticulum (SR) Ca2+ -ATPase type-2a (SERCA2a; 1:2,000, sc-8095, Santa Cruz Biotechnology), transforming growth factor β1 (TGF-β1, 1:1000, ab9758, Abcam), and vimentin (1:1,000, sc373717, Santa Cruz Biotechnology) were determined using appropriate primary antibodies.

    Techniques: Western Blot, Expressing, CTL Assay

    Gene expression analysis. Using a customized NanoString's nCounter® Elements TagSet panel of 27 genes coding for proteins regulated in hypertrophy/heart failure, including Ca 2+ and K + handling proteins, steady state mRNA-concentrations were analyzed in 22 days old EHTs after AAV6-mediated overexpression of S100A4 ( n = 6) under the control of the human cardiac troponin T promoter ( TNNT2) or EHTs after treatment with an empty vector ( n = 5). Data were analyzed with nCounter® Sprint Profiler including background subtraction using negative controls and normalization to 6 housekeeping genes ( ABCF1, ACTB, CLTC, GAPDH, PGK1, TUBB ). Data represented the mean of normalized counts and were expressed as fold-change in EHTs overexpressing S100A4V5 vs. control EHTs. A threshold of −0.2- and +0.25-fold difference related to empty vector was considered a relevant result and such genes were marked in bold.

    Journal: Frontiers in Physiology

    Article Title: S100A4 as a Target of the E3-Ligase Asb2β and Its Effect on Engineered Heart Tissue

    doi: 10.3389/fphys.2018.01292

    Figure Lengend Snippet: Gene expression analysis. Using a customized NanoString's nCounter® Elements TagSet panel of 27 genes coding for proteins regulated in hypertrophy/heart failure, including Ca 2+ and K + handling proteins, steady state mRNA-concentrations were analyzed in 22 days old EHTs after AAV6-mediated overexpression of S100A4 ( n = 6) under the control of the human cardiac troponin T promoter ( TNNT2) or EHTs after treatment with an empty vector ( n = 5). Data were analyzed with nCounter® Sprint Profiler including background subtraction using negative controls and normalization to 6 housekeeping genes ( ABCF1, ACTB, CLTC, GAPDH, PGK1, TUBB ). Data represented the mean of normalized counts and were expressed as fold-change in EHTs overexpressing S100A4V5 vs. control EHTs. A threshold of −0.2- and +0.25-fold difference related to empty vector was considered a relevant result and such genes were marked in bold.

    Article Snippet: Membranes were blocked in 5% milk and incubated with antibodies against S100A4 (anti-S100A4 rabbit-polyclonal, Dako A5114, 1:500), ß-actin (anti-ß-actin mouse-monoclonal, 1:20,000), GAPDH (anti-GAPDH mouse-monoclonal, HyTest 5G4-6C5, 1:5,000), FLAG (anti-FLAG mouse-monoclonal, Sigma F3165, 1:5,000), V5 (anti-V5 mouse-polyclonal, Invitrogen PA1-993, 1:1,000), GFP (anti-GFP rabbit-polyclonal, Abcam ab6556, 1:2,000), and total ERK (anti-Total ERK rabbit-polyclonal, Cell Signaling #9102, 1:2,000).

    Techniques: Expressing, Over Expression, Plasmid Preparation

    Glucocorticoids suppress Wnt16 in vitro . ( A – C ) Bone marrow stromal cells derived from wildtype mice were differentiated towards osteoblasts for 7 days and treated with ( A ) various concentrations of dexamethasone (DEX) and ( B ) various durations. N = 4–12. Wnt16 gene expression was quantified using real-time PCR. Beta-actin was used as housekeeping gene. ( C ) Cells were treated with 100 nM DEX or 48 h. Protein levels of Wnt16 and Gapdh were detected using Western blot. Two independent experiments are shown out of four. Each Western blot represents an own gel. Full gels are shown in Suppl. Fig. 1 . ( D ) Calvarial bone from 3–5 days old C57BL/6 pups were cultured for 24 h in the presence or absence of 1 µM DEX. Wnt16 gene expression was quantified using real-time PCR (n = 6); protein expression using Western blot (n = 4). Numbers indicate semi-quantification of Western blots. Each Western blot represents an own gel. Full gels are shown in Suppl. Fig. 1 . *p

    Journal: Scientific Reports

    Article Title: Glucocorticoids suppress Wnt16 expression in osteoblasts in vitro and in vivo

    doi: 10.1038/s41598-018-26300-z

    Figure Lengend Snippet: Glucocorticoids suppress Wnt16 in vitro . ( A – C ) Bone marrow stromal cells derived from wildtype mice were differentiated towards osteoblasts for 7 days and treated with ( A ) various concentrations of dexamethasone (DEX) and ( B ) various durations. N = 4–12. Wnt16 gene expression was quantified using real-time PCR. Beta-actin was used as housekeeping gene. ( C ) Cells were treated with 100 nM DEX or 48 h. Protein levels of Wnt16 and Gapdh were detected using Western blot. Two independent experiments are shown out of four. Each Western blot represents an own gel. Full gels are shown in Suppl. Fig. 1 . ( D ) Calvarial bone from 3–5 days old C57BL/6 pups were cultured for 24 h in the presence or absence of 1 µM DEX. Wnt16 gene expression was quantified using real-time PCR (n = 6); protein expression using Western blot (n = 4). Numbers indicate semi-quantification of Western blots. Each Western blot represents an own gel. Full gels are shown in Suppl. Fig. 1 . *p

    Article Snippet: After blocking for 1 h with 5% milk powder in Tris-buffered saline with 1% Tween-20 (TBS-T), membranes were incubated with antibodies against Wnt16 (1:500, Abcam, #ab109437) and GAPDH (1:2,000, Hytest, #5G4) overnight.

    Techniques: In Vitro, Derivative Assay, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Cell Culture

    Everolimus (EV) inhibits cancer cell growth in vitro and in vivo. a The murine melanoma cell line B16-F10 and the human breast cancer cell lines MCF-7 and MDA-MB-231 were treated with EV in a dose-dependent (0, 1, 10, and 100 nM) and time-dependent (0, 24, 48, and 72 h) manner. Cell viability was assessed with the CellTiter-Blue® assay. b Western blots used to assess the ability of EV concentrations to inhibit the phosphorylation of the mammalian target of rapamycin (mTOR) protein and p70 S6 kinase after 24 h of treatment in the cell lines investigated. Glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) is shown as the housekeeping control. c Female immunocompetent (C57BL/6) and immunocompromised (NMRI nude) mice were inoculated subcutaneously with B16-F10 and MDA-MB-231 cells, respectively. Tumor growth was assessed after daily treatment with 1 mg/kg of EV for 2 and 4 weeks in each respective model. In vitro and in vivo data are shown as mean ± SD of at least three independent experiments or ten mice per group, respectively. Cell viability assays were analyzed for each time point using two-way analysis of variance and in vivo data by Student’s t test. Significance between EV treatments and the control condition was apparent only at 72 h and is indicated by asterisks on the graphs. In the B16-F10 graph, EV treatment was significant at inhibiting viability only at 72 h at concentrations of 10 and 100 nM. In the MDA-MB-231 and MCF-7 graphs, all concentrations of EV were significant to the same degree. ** p

    Journal: Breast Cancer Research : BCR

    Article Title: Concurrent antitumor and bone-protective effects of everolimus in osteotropic breast cancer

    doi: 10.1186/s13058-017-0885-7

    Figure Lengend Snippet: Everolimus (EV) inhibits cancer cell growth in vitro and in vivo. a The murine melanoma cell line B16-F10 and the human breast cancer cell lines MCF-7 and MDA-MB-231 were treated with EV in a dose-dependent (0, 1, 10, and 100 nM) and time-dependent (0, 24, 48, and 72 h) manner. Cell viability was assessed with the CellTiter-Blue® assay. b Western blots used to assess the ability of EV concentrations to inhibit the phosphorylation of the mammalian target of rapamycin (mTOR) protein and p70 S6 kinase after 24 h of treatment in the cell lines investigated. Glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) is shown as the housekeeping control. c Female immunocompetent (C57BL/6) and immunocompromised (NMRI nude) mice were inoculated subcutaneously with B16-F10 and MDA-MB-231 cells, respectively. Tumor growth was assessed after daily treatment with 1 mg/kg of EV for 2 and 4 weeks in each respective model. In vitro and in vivo data are shown as mean ± SD of at least three independent experiments or ten mice per group, respectively. Cell viability assays were analyzed for each time point using two-way analysis of variance and in vivo data by Student’s t test. Significance between EV treatments and the control condition was apparent only at 72 h and is indicated by asterisks on the graphs. In the B16-F10 graph, EV treatment was significant at inhibiting viability only at 72 h at concentrations of 10 and 100 nM. In the MDA-MB-231 and MCF-7 graphs, all concentrations of EV were significant to the same degree. ** p

    Article Snippet: An antibody for the housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (5G4) was purchased from HyTest Oy (Turku, Finland).

    Techniques: In Vitro, In Vivo, Multiple Displacement Amplification, CtB Assay, Western Blot, Mouse Assay