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Cayman Chemical doxorubicin hydrochloride
Doxorubicin Hydrochloride, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/doxorubicin hydrochloride/product/Cayman Chemical
Average 93 stars, based on 4 article reviews
Price from $9.99 to $1999.99
doxorubicin hydrochloride - by Bioz Stars, 2020-01
93/100 stars

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In Vivo:

Article Title: Antitumor profile of the PI3K inhibitor ZSTK474 in human sarcoma cell lines
Article Snippet: ZSTK474, ZSTK534, ZSTK778, ZSTK1741 and ZSTK2209 were kindly gifted from Zenyaku Kogyo Co., Ltd. For in vivo studies, a solid dispersion form of ZSTK474 was suspended with distilled water (Otsuka Pharmaceutical Factory, Inc., Tokushima, Japan). .. Doxorubicin was purchased from Cayman Chemical Company (Ann Arbor, MI) and dissolved with saline (Otsuka Pharmaceutical Factory, Inc., Tokushima, Japan).

Incubation:

Article Title: CBX Chromodomain Inhibition Enhances Chemotherapy Response in Glioblastoma Multiforme
Article Snippet: Cells were dosed in a grid format for every combination of drug at the designated dose: PRT4165 40 µM (Cayman), PTC209 200 nM (Cayman), DZnep 25 µM (Cayman), GSK343 400 nM (Cayman), MS37452 200 µM (Cayman), Doxorubicin 200 nM, temozolomide 50 µM (Cayman), SAHA 1 µM (Cayman). .. Following five days of treatment, cells were fixed with 50 percent trichloroacetic acid (TCA) for an hour at 4 °C, washed with water, incubated in sulforhodamine B for 10 minutes, washed with 1 percent acetic acid and dried overnight.

Article Title: The prionlike domain of FUS is multiphosphorylated following DNA damage without altering nuclear localization
Article Snippet: Cells were lysed by resuspension and incubation in lysis buffer (200 mM NaCl,100 mM Tris-HCl, 0.5% sodium deoxycholate, 1% Triton X-100, 0.2% SDS, 660 mM phenylmethylsulfonyl fluoride, 100 μl protease inhibitor cocktail (Sigma P8215), 100 μl phosphatase inhibitor cocktail (Sigma P0044), 1250 U Benzonase nuclease) while on ice. .. The following drugs were used to treat the human cell lines: calicheamicin-γ (a generous gift from Pfizer), calyculin-A (Sigma C5552), camptothecin (Sigma C9911), etoposide (Sigma E1383), doxorubicin (Cayman Chemical 15007), okadaic acid (Sigma O9381), bleomycin (Cayman Chemical 13877), DMSO (Sigma D8418), DNA-PK inhibitor (Selleckchem NU7441), sorbitol (Sigma A1876), and Adox (Sigma A7154).

Article Title: Follicular lymphoma–associated mutations in vacuolar ATPase ATP6V1B2 activate autophagic flux and mTOR
Article Snippet: MRT 68921 (catalog 1190379), bafilomycin A1 (catalog 11038), rapamycin (catalog 13346), torin 1 (catalog 10997), doxorubicin (catalog 15007), dexamethasone (catalog 11015), vincristine (catalog 11764), and chloroquine (catalog 14194), all procured from Cayman Chemical, were diluted serially in medium, and DMSO alone was used as a vehicle control. .. After 72 hours of incubation, cell viability was measured with CellTiter-Glo (Promega, G7572) and quantified on a GloMax microplate reader (Promega, SA3030).

Proliferation Assay:

Article Title: WDR23 regulates NRF2 independently of KEAP1
Article Snippet: Forty-eight hours post-transfection, cells were treated with the indicated chemical: Etoposide (Cayman), Doxorubicin (Cayman), or Cisplatin (Cayman). .. For viability assays, after forty-eight hours of treatment, cells were assayed using the Vybrant MTT Cell Proliferation Assay Kit (Thermo Scientific), performed according to the manufacturer’s protocol.

Infection:

Article Title: Lsd1 as a therapeutic target in Gfi1-activated medulloblastoma
Article Snippet: Cells were treated with vehicle (DMSO), 0.1 or 0.5 µM doxorubicin (Cayman Chemical cat #15007), or they were irradiated with 0, 1, 2, 4, or 8 Gy using a low-dose cesium 137 irradiator at the Sanford Burnham Prebys Animal Facility. .. DNp53-expressing MG tumors were generated by isolating MG tumor cells, infecting with MSCV-DNp53-IRES-GFP viruses, sorting for infected cells and retransplanting the cells into new NSG mice.

Article Title: Ewing sarcoma resistance to SP-2509 is not mediated through KDM1A/LSD1 mutation
Article Snippet: Cell culture supernatants were tested yearly for Mycoplasma infection using a PCR based detection kit (Southern Biotech, USA) with cells authenticated by STR profiling (Genetica LabCorp, USA). .. SP-2509 was provided by Dr Sunil Sharma (Huntsman Cancer Institute, Utah), Doxorubicin hydrochloride, Etoposide, Vincristine sulfate, and Vorinostat/SAHA were purchased from Cayman Chemical, and Entinostat was purchased from Selleckchem.

Expressing:

Article Title: Long-lasting reduction in clonogenic potential of colorectal cancer cells by sequential treatments with 5-azanucleosides and topoisomerase inhibitors
Article Snippet: After 2 days, the culture medium was changed and the cells were treated either with 5-aza-dC or 5-aza-C along either with irinotecan (5–75 μM; Cayman Chemical), etoposide (5–50 μM; Sigma-Aldrich), doxorubicin (0.05–0.9 μM; Cayman Chemical), or mitoxantrone (0.05–1 μM; Cayman Chemical). .. In Western blotting analyses of Akt, phospho-Akt, mTOR, phospho-mTOR, p70S6K, and phospho-p70S6K expression, the cells were treated with 5-aza-dC (0.1–1 μM) 1 day post-seeding.

Planar Chromatography:

Article Title: Allele-specific silencing of mutant p53 attenuates dominant-negative and gain-of-function activities
Article Snippet: HCT116wt/R248W and SW48wt/R273H cell lines, which were genetically engineered to have a heterozygous mutation at the endogenous p53 locus, were purchased from Horizon Discovery Group plc (Cambridge, United Kingdom). .. When required, cells were treated with 5 μM of a MDM2 inhibitor Nutlin-3a (Sigma-Aldrich, St. Louis, MO) or doxorubicin at different concentrations (Cayman Chemical, Ann Arbor, Michigan) for 24 to 48h to activate p53 [ , ].

Modification:

Article Title: Allele-specific silencing of mutant p53 attenuates dominant-negative and gain-of-function activities
Article Snippet: Cell lines All of the following human cell lines (with different p53 status) were maintained in Dulbecco's Modified Eagle's Medium (DMEM) or Roswell Park Memorial Institute (RPMI) medium with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin: human osteosarcoma KHOS/NP (p53R156P ), U2OS (p53wt ), colorectal carcinoma HCT116 (p53wt ), colorectal adenocarcinoma SW48 (p53wt ), tongue squamous cell carcinoma CAL33 (p53R175H ), pancreatic carcinoma MiaPaCa2 (p53R248W ), colorectal adenocarcinoma SW620 (p53R273H ), and mouse osteosarcoma 318-1 (p53R172H ) as described in [ ]. .. When required, cells were treated with 5 μM of a MDM2 inhibitor Nutlin-3a (Sigma-Aldrich, St. Louis, MO) or doxorubicin at different concentrations (Cayman Chemical, Ann Arbor, Michigan) for 24 to 48h to activate p53 [ , ].

Western Blot:

Article Title: Selective in vivo metabolic cell-labeling-mediated cancer targeting
Article Snippet: EZ-Link phosphine–PEG3 –biotin, streptavidin–horseradish peroxidase (HRP), and Pierce ECL western blotting substrate were purchased from Thermo Fisher Scientific (Waltham, MA, USA). .. Doxorubicin hydrochloride was purchased from Cayman Chemical Company (Ann Arbor, MI, USA).

Article Title: Lsd1 as a therapeutic target in Gfi1-activated medulloblastoma
Article Snippet: Cells were treated with vehicle (DMSO), 0.1 or 0.5 µM doxorubicin (Cayman Chemical cat #15007), or they were irradiated with 0, 1, 2, 4, or 8 Gy using a low-dose cesium 137 irradiator at the Sanford Burnham Prebys Animal Facility. .. Samples were collected 4 hrs after treatment or irradiation for analysis of p53 and p21 protein levels by Western blot.

Article Title: Long-lasting reduction in clonogenic potential of colorectal cancer cells by sequential treatments with 5-azanucleosides and topoisomerase inhibitors
Article Snippet: The cells were seeded at densities of 4 × 103 cells/0.1 ml (0.32 cm2 ) (cell viability assay), 6 × 103 –1 × 104 cells/0.3 ml (0.7 cm2 ) (microscopic images), 2 × 104 cells/0.5 ml (1.9 cm2 ) (flow cytometry), 1 × 105 cells/3 ml (9.5 cm2 ) (long-term colony formation assay, serial replating assay), 3.5 × 105 cells/3–4 ml (21 cm2 ) (Western blotting). .. After 2 days, the culture medium was changed and the cells were treated either with 5-aza-dC or 5-aza-C along either with irinotecan (5–75 μM; Cayman Chemical), etoposide (5–50 μM; Sigma-Aldrich), doxorubicin (0.05–0.9 μM; Cayman Chemical), or mitoxantrone (0.05–1 μM; Cayman Chemical).

Flow Cytometry:

Article Title: Long-lasting reduction in clonogenic potential of colorectal cancer cells by sequential treatments with 5-azanucleosides and topoisomerase inhibitors
Article Snippet: The cells were seeded at densities of 4 × 103 cells/0.1 ml (0.32 cm2 ) (cell viability assay), 6 × 103 –1 × 104 cells/0.3 ml (0.7 cm2 ) (microscopic images), 2 × 104 cells/0.5 ml (1.9 cm2 ) (flow cytometry), 1 × 105 cells/3 ml (9.5 cm2 ) (long-term colony formation assay, serial replating assay), 3.5 × 105 cells/3–4 ml (21 cm2 ) (Western blotting). .. After 2 days, the culture medium was changed and the cells were treated either with 5-aza-dC or 5-aza-C along either with irinotecan (5–75 μM; Cayman Chemical), etoposide (5–50 μM; Sigma-Aldrich), doxorubicin (0.05–0.9 μM; Cayman Chemical), or mitoxantrone (0.05–1 μM; Cayman Chemical).

Protease Inhibitor:

Article Title: The prionlike domain of FUS is multiphosphorylated following DNA damage without altering nuclear localization
Article Snippet: Cells were lysed by resuspension and incubation in lysis buffer (200 mM NaCl,100 mM Tris-HCl, 0.5% sodium deoxycholate, 1% Triton X-100, 0.2% SDS, 660 mM phenylmethylsulfonyl fluoride, 100 μl protease inhibitor cocktail (Sigma P8215), 100 μl phosphatase inhibitor cocktail (Sigma P0044), 1250 U Benzonase nuclease) while on ice. .. The following drugs were used to treat the human cell lines: calicheamicin-γ (a generous gift from Pfizer), calyculin-A (Sigma C5552), camptothecin (Sigma C9911), etoposide (Sigma E1383), doxorubicin (Cayman Chemical 15007), okadaic acid (Sigma O9381), bleomycin (Cayman Chemical 13877), DMSO (Sigma D8418), DNA-PK inhibitor (Selleckchem NU7441), sorbitol (Sigma A1876), and Adox (Sigma A7154).

Cell Culture:

Article Title: The prionlike domain of FUS is multiphosphorylated following DNA damage without altering nuclear localization
Article Snippet: Paragraph title: Cell culture ... The following drugs were used to treat the human cell lines: calicheamicin-γ (a generous gift from Pfizer), calyculin-A (Sigma C5552), camptothecin (Sigma C9911), etoposide (Sigma E1383), doxorubicin (Cayman Chemical 15007), okadaic acid (Sigma O9381), bleomycin (Cayman Chemical 13877), DMSO (Sigma D8418), DNA-PK inhibitor (Selleckchem NU7441), sorbitol (Sigma A1876), and Adox (Sigma A7154).

Article Title: Long-lasting reduction in clonogenic potential of colorectal cancer cells by sequential treatments with 5-azanucleosides and topoisomerase inhibitors
Article Snippet: Paragraph title: Cell culture and treatment ... After 2 days, the culture medium was changed and the cells were treated either with 5-aza-dC or 5-aza-C along either with irinotecan (5–75 μM; Cayman Chemical), etoposide (5–50 μM; Sigma-Aldrich), doxorubicin (0.05–0.9 μM; Cayman Chemical), or mitoxantrone (0.05–1 μM; Cayman Chemical).

Article Title: Follicular lymphoma–associated mutations in vacuolar ATPase ATP6V1B2 activate autophagic flux and mTOR
Article Snippet: Purified FL cells were plated in RPMI 1640 medium supplemented with 20% heat-inactivated FBS at a density of 106 cells/ml and cultured at 37°C with 5% CO2 . .. MRT 68921 (catalog 1190379), bafilomycin A1 (catalog 11038), rapamycin (catalog 13346), torin 1 (catalog 10997), doxorubicin (catalog 15007), dexamethasone (catalog 11015), vincristine (catalog 11764), and chloroquine (catalog 14194), all procured from Cayman Chemical, were diluted serially in medium, and DMSO alone was used as a vehicle control.

Article Title: Ewing sarcoma resistance to SP-2509 is not mediated through KDM1A/LSD1 mutation
Article Snippet: Paragraph title: Reagents and cell culture ... SP-2509 was provided by Dr Sunil Sharma (Huntsman Cancer Institute, Utah), Doxorubicin hydrochloride, Etoposide, Vincristine sulfate, and Vorinostat/SAHA were purchased from Cayman Chemical, and Entinostat was purchased from Selleckchem.

Article Title: Microfluidics 3D gel-island chip for single cell isolation and lineage-dependent drug responses study
Article Snippet: For the demonstration of anticancer drug susceptibility testing, T47D Notch+ cells were loaded with collagen gel at a concentration of 2 mg/mL and cultured in gel-islands for 48 hours, as described above. .. Then, cells were treated with 0.3 μM Doxorubicin (Cayman Chemical 15007) and 50 μM Cisplatin (Cayman Chemical 13119) for 72 hours.

Generated:

Article Title: Lsd1 as a therapeutic target in Gfi1-activated medulloblastoma
Article Snippet: Cells were treated with vehicle (DMSO), 0.1 or 0.5 µM doxorubicin (Cayman Chemical cat #15007), or they were irradiated with 0, 1, 2, 4, or 8 Gy using a low-dose cesium 137 irradiator at the Sanford Burnham Prebys Animal Facility. .. DNp53-expressing MG tumors were generated by isolating MG tumor cells, infecting with MSCV-DNp53-IRES-GFP viruses, sorting for infected cells and retransplanting the cells into new NSG mice.

Imaging:

Article Title: Microfluidics 3D gel-island chip for single cell isolation and lineage-dependent drug responses study
Article Snippet: Then, cells were treated with 0.3 μM Doxorubicin (Cayman Chemical 15007) and 50 μM Cisplatin (Cayman Chemical 13119) for 72 hours. .. To determine the cell viability after drug treatment, cells were stained using the Live/Dead Viability/Cytotoxicity Kit for mammalian cells (Life Technologies, L-3224) and were put in an incubator (37°C, 5% CO2 ) for 30 minutes, followed by fluorescence microscopy imaging.

Polymerase Chain Reaction:

Article Title: Ewing sarcoma resistance to SP-2509 is not mediated through KDM1A/LSD1 mutation
Article Snippet: Cell culture supernatants were tested yearly for Mycoplasma infection using a PCR based detection kit (Southern Biotech, USA) with cells authenticated by STR profiling (Genetica LabCorp, USA). .. SP-2509 was provided by Dr Sunil Sharma (Huntsman Cancer Institute, Utah), Doxorubicin hydrochloride, Etoposide, Vincristine sulfate, and Vorinostat/SAHA were purchased from Cayman Chemical, and Entinostat was purchased from Selleckchem.

Molecular Weight:

Article Title: Selective in vivo metabolic cell-labeling-mediated cancer targeting
Article Snippet: Doxorubicin hydrochloride was purchased from Cayman Chemical Company (Ann Arbor, MI, USA). .. HOOC–PEG–NH2 (1 k molecular weight) was purchased from Nanocs Inc. (New York, NY, USA).

MTT Assay:

Article Title: WDR23 regulates NRF2 independently of KEAP1
Article Snippet: Forty-eight hours post-transfection, cells were treated with the indicated chemical: Etoposide (Cayman), Doxorubicin (Cayman), or Cisplatin (Cayman). .. For viability assays, after forty-eight hours of treatment, cells were assayed using the Vybrant MTT Cell Proliferation Assay Kit (Thermo Scientific), performed according to the manufacturer’s protocol.

Digital Holographic Microscopy:

Article Title: Quantifying Drug-Induced Nanomechanics and Mechanical Effects to Single Cardiomyocytes for Optimal Drug Administration To Minimize Cardiotoxicity
Article Snippet: The cases treated by one type of drug include the doxorubicin (Cayman Chemical Co.) treated group and the dexrazoxane (Sigma-Aldrich) treated group. .. The cardiomyocytes were continuously monitored by DHM or AFM for about 2 h. The temperature inside the fluidic chamber was maintained at 37 °C.

Fluorescence:

Article Title: Microfluidics 3D gel-island chip for single cell isolation and lineage-dependent drug responses study
Article Snippet: Then, cells were treated with 0.3 μM Doxorubicin (Cayman Chemical 15007) and 50 μM Cisplatin (Cayman Chemical 13119) for 72 hours. .. To determine the cell viability after drug treatment, cells were stained using the Live/Dead Viability/Cytotoxicity Kit for mammalian cells (Life Technologies, L-3224) and were put in an incubator (37°C, 5% CO2 ) for 30 minutes, followed by fluorescence microscopy imaging.

Mutagenesis:

Article Title: Allele-specific silencing of mutant p53 attenuates dominant-negative and gain-of-function activities
Article Snippet: HCT116wt/R248W and SW48wt/R273H cell lines, which were genetically engineered to have a heterozygous mutation at the endogenous p53 locus, were purchased from Horizon Discovery Group plc (Cambridge, United Kingdom). .. When required, cells were treated with 5 μM of a MDM2 inhibitor Nutlin-3a (Sigma-Aldrich, St. Louis, MO) or doxorubicin at different concentrations (Cayman Chemical, Ann Arbor, Michigan) for 24 to 48h to activate p53 [ , ].

Isolation:

Article Title: Quantifying Drug-Induced Nanomechanics and Mechanical Effects to Single Cardiomyocytes for Optimal Drug Administration To Minimize Cardiotoxicity
Article Snippet: Primary mouse cardiomyocytes were isolated from wild-type mice and used as samples in the experiments (detailed isolation procedures are described in the ). .. The cases treated by one type of drug include the doxorubicin (Cayman Chemical Co.) treated group and the dexrazoxane (Sigma-Aldrich) treated group.

Article Title: Lsd1 as a therapeutic target in Gfi1-activated medulloblastoma
Article Snippet: Induction of p53 DNA damage response MEFs were isolated from C57BL/6 embryos at embryonic day 14.5. .. Cells were treated with vehicle (DMSO), 0.1 or 0.5 µM doxorubicin (Cayman Chemical cat #15007), or they were irradiated with 0, 1, 2, 4, or 8 Gy using a low-dose cesium 137 irradiator at the Sanford Burnham Prebys Animal Facility.

Transfection:

Article Title: WDR23 regulates NRF2 independently of KEAP1
Article Snippet: Chemotherapy treatment and cytology HEK-293T cells were transiently transfected with indicated plasmids. .. Forty-eight hours post-transfection, cells were treated with the indicated chemical: Etoposide (Cayman), Doxorubicin (Cayman), or Cisplatin (Cayman).

Microscopy:

Article Title: Microfluidics 3D gel-island chip for single cell isolation and lineage-dependent drug responses study
Article Snippet: Then, cells were treated with 0.3 μM Doxorubicin (Cayman Chemical 15007) and 50 μM Cisplatin (Cayman Chemical 13119) for 72 hours. .. To determine the cell viability after drug treatment, cells were stained using the Live/Dead Viability/Cytotoxicity Kit for mammalian cells (Life Technologies, L-3224) and were put in an incubator (37°C, 5% CO2 ) for 30 minutes, followed by fluorescence microscopy imaging.

Mouse Assay:

Article Title: Quantifying Drug-Induced Nanomechanics and Mechanical Effects to Single Cardiomyocytes for Optimal Drug Administration To Minimize Cardiotoxicity
Article Snippet: Primary mouse cardiomyocytes were isolated from wild-type mice and used as samples in the experiments (detailed isolation procedures are described in the ). .. The cases treated by one type of drug include the doxorubicin (Cayman Chemical Co.) treated group and the dexrazoxane (Sigma-Aldrich) treated group.

Article Title: Lsd1 as a therapeutic target in Gfi1-activated medulloblastoma
Article Snippet: Cells were treated with vehicle (DMSO), 0.1 or 0.5 µM doxorubicin (Cayman Chemical cat #15007), or they were irradiated with 0, 1, 2, 4, or 8 Gy using a low-dose cesium 137 irradiator at the Sanford Burnham Prebys Animal Facility. .. DNp53-expressing MG tumors were generated by isolating MG tumor cells, infecting with MSCV-DNp53-IRES-GFP viruses, sorting for infected cells and retransplanting the cells into new NSG mice.

Cytometry:

Article Title: Long-lasting reduction in clonogenic potential of colorectal cancer cells by sequential treatments with 5-azanucleosides and topoisomerase inhibitors
Article Snippet: The cells were seeded at densities of 4 × 103 cells/0.1 ml (0.32 cm2 ) (cell viability assay), 6 × 103 –1 × 104 cells/0.3 ml (0.7 cm2 ) (microscopic images), 2 × 104 cells/0.5 ml (1.9 cm2 ) (flow cytometry), 1 × 105 cells/3 ml (9.5 cm2 ) (long-term colony formation assay, serial replating assay), 3.5 × 105 cells/3–4 ml (21 cm2 ) (Western blotting). .. After 2 days, the culture medium was changed and the cells were treated either with 5-aza-dC or 5-aza-C along either with irinotecan (5–75 μM; Cayman Chemical), etoposide (5–50 μM; Sigma-Aldrich), doxorubicin (0.05–0.9 μM; Cayman Chemical), or mitoxantrone (0.05–1 μM; Cayman Chemical).

Lysis:

Article Title: The prionlike domain of FUS is multiphosphorylated following DNA damage without altering nuclear localization
Article Snippet: Cells were lysed by resuspension and incubation in lysis buffer (200 mM NaCl,100 mM Tris-HCl, 0.5% sodium deoxycholate, 1% Triton X-100, 0.2% SDS, 660 mM phenylmethylsulfonyl fluoride, 100 μl protease inhibitor cocktail (Sigma P8215), 100 μl phosphatase inhibitor cocktail (Sigma P0044), 1250 U Benzonase nuclease) while on ice. .. The following drugs were used to treat the human cell lines: calicheamicin-γ (a generous gift from Pfizer), calyculin-A (Sigma C5552), camptothecin (Sigma C9911), etoposide (Sigma E1383), doxorubicin (Cayman Chemical 15007), okadaic acid (Sigma O9381), bleomycin (Cayman Chemical 13877), DMSO (Sigma D8418), DNA-PK inhibitor (Selleckchem NU7441), sorbitol (Sigma A1876), and Adox (Sigma A7154).

Purification:

Article Title: Follicular lymphoma–associated mutations in vacuolar ATPase ATP6V1B2 activate autophagic flux and mTOR
Article Snippet: Purified FL cells were plated in RPMI 1640 medium supplemented with 20% heat-inactivated FBS at a density of 106 cells/ml and cultured at 37°C with 5% CO2 . .. MRT 68921 (catalog 1190379), bafilomycin A1 (catalog 11038), rapamycin (catalog 13346), torin 1 (catalog 10997), doxorubicin (catalog 15007), dexamethasone (catalog 11015), vincristine (catalog 11764), and chloroquine (catalog 14194), all procured from Cayman Chemical, were diluted serially in medium, and DMSO alone was used as a vehicle control.

Viability Assay:

Article Title: Long-lasting reduction in clonogenic potential of colorectal cancer cells by sequential treatments with 5-azanucleosides and topoisomerase inhibitors
Article Snippet: The cells were seeded at densities of 4 × 103 cells/0.1 ml (0.32 cm2 ) (cell viability assay), 6 × 103 –1 × 104 cells/0.3 ml (0.7 cm2 ) (microscopic images), 2 × 104 cells/0.5 ml (1.9 cm2 ) (flow cytometry), 1 × 105 cells/3 ml (9.5 cm2 ) (long-term colony formation assay, serial replating assay), 3.5 × 105 cells/3–4 ml (21 cm2 ) (Western blotting). .. After 2 days, the culture medium was changed and the cells were treated either with 5-aza-dC or 5-aza-C along either with irinotecan (5–75 μM; Cayman Chemical), etoposide (5–50 μM; Sigma-Aldrich), doxorubicin (0.05–0.9 μM; Cayman Chemical), or mitoxantrone (0.05–1 μM; Cayman Chemical).

Irradiation:

Article Title: Lsd1 as a therapeutic target in Gfi1-activated medulloblastoma
Article Snippet: .. Cells were treated with vehicle (DMSO), 0.1 or 0.5 µM doxorubicin (Cayman Chemical cat #15007), or they were irradiated with 0, 1, 2, 4, or 8 Gy using a low-dose cesium 137 irradiator at the Sanford Burnham Prebys Animal Facility. .. Samples were collected 4 hrs after treatment or irradiation for analysis of p53 and p21 protein levels by Western blot.

Article Title: The prionlike domain of FUS is multiphosphorylated following DNA damage without altering nuclear localization
Article Snippet: The following drugs were used to treat the human cell lines: calicheamicin-γ (a generous gift from Pfizer), calyculin-A (Sigma C5552), camptothecin (Sigma C9911), etoposide (Sigma E1383), doxorubicin (Cayman Chemical 15007), okadaic acid (Sigma O9381), bleomycin (Cayman Chemical 13877), DMSO (Sigma D8418), DNA-PK inhibitor (Selleckchem NU7441), sorbitol (Sigma A1876), and Adox (Sigma A7154). .. UV irradiation was carried out by a standard germicidal ultraviolet light with 95% of the ultraviolet radiation in the 254-nm region at 37°C.

In Vitro:

Article Title: Antitumor profile of the PI3K inhibitor ZSTK474 in human sarcoma cell lines
Article Snippet: Doxorubicin was purchased from Cayman Chemical Company (Ann Arbor, MI) and dissolved with saline (Otsuka Pharmaceutical Factory, Inc., Tokushima, Japan). .. Pazopanib was purchased from Cellagen Technology (San Diego, CA), dissolved with DMSO for in vitro studies, and suspended with 5% hydroxypropylmethyl cellulose (HPMC) for in vivo studies.

Colony Assay:

Article Title: Long-lasting reduction in clonogenic potential of colorectal cancer cells by sequential treatments with 5-azanucleosides and topoisomerase inhibitors
Article Snippet: The cells were seeded at densities of 4 × 103 cells/0.1 ml (0.32 cm2 ) (cell viability assay), 6 × 103 –1 × 104 cells/0.3 ml (0.7 cm2 ) (microscopic images), 2 × 104 cells/0.5 ml (1.9 cm2 ) (flow cytometry), 1 × 105 cells/3 ml (9.5 cm2 ) (long-term colony formation assay, serial replating assay), 3.5 × 105 cells/3–4 ml (21 cm2 ) (Western blotting). .. After 2 days, the culture medium was changed and the cells were treated either with 5-aza-dC or 5-aza-C along either with irinotecan (5–75 μM; Cayman Chemical), etoposide (5–50 μM; Sigma-Aldrich), doxorubicin (0.05–0.9 μM; Cayman Chemical), or mitoxantrone (0.05–1 μM; Cayman Chemical).

Concentration Assay:

Article Title: Microfluidics 3D gel-island chip for single cell isolation and lineage-dependent drug responses study
Article Snippet: For the demonstration of anticancer drug susceptibility testing, T47D Notch+ cells were loaded with collagen gel at a concentration of 2 mg/mL and cultured in gel-islands for 48 hours, as described above. .. Then, cells were treated with 0.3 μM Doxorubicin (Cayman Chemical 15007) and 50 μM Cisplatin (Cayman Chemical 13119) for 72 hours.

Staining:

Article Title: Microfluidics 3D gel-island chip for single cell isolation and lineage-dependent drug responses study
Article Snippet: Then, cells were treated with 0.3 μM Doxorubicin (Cayman Chemical 15007) and 50 μM Cisplatin (Cayman Chemical 13119) for 72 hours. .. To determine the cell viability after drug treatment, cells were stained using the Live/Dead Viability/Cytotoxicity Kit for mammalian cells (Life Technologies, L-3224) and were put in an incubator (37°C, 5% CO2 ) for 30 minutes, followed by fluorescence microscopy imaging.

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  • 96
    Cayman Chemical doxorubicin
    Effect of <t>doxorubicin</t> and DoxQ on adult rat cardiomyocyte viability. a Dox versus DoxQ dose response. N = 3; mean ± SEM. * P
    Doxorubicin, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 96/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/doxorubicin/product/Cayman Chemical
    Average 96 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    doxorubicin - by Bioz Stars, 2020-01
    96/100 stars
      Buy from Supplier

    78
    Cayman Chemical cardiomyocyte nuclear nf κb activity
    Effects of dobutamine (DOB) and lipopolysaccharide (LPS) on <t>cardiomyocyte</t> Bcl-2 levels, mitochondrial Bax contents, mitochondrial membrane potential and cytochrome c release. Adult mouse ventricular myocytes were treated with DOB (0.02 μM), LPS (10 ng/mL), their combination, or vehicle for 12 hours. (A) Bcl-2 levels were analyzed by western blotting of whole cell lysates. (B) Mitochondrial Bax protein contents were detected by western blotting of mitochondrial proteins and voltage-dependent anion channel (VDAC) was immunoblotted as a mitochondrial marker. (C) Confocal images of JC-1 fluorescence. Mitochondrial membrane potential was visualized in ventricular myocytes stained with JC-1, an indicator of mitochondrial function, green fluorescence indicates monomeric JC-1 (left), red fluorescence represents aggregate JC-1 (middle) and merged channels are shown (right): scale bar = 20 μm. (D) The ratio of monomeric and aggregated JC-1, indicating changes in mitochondrial membrane potential. (E) Cytosolic cytochrome c (Cyt c) contents were analyzed by western blotting of cytosolic proteins and GAPDH was used as an internal control: mean ± standard error of the mean (n = 3). * P
    Cardiomyocyte Nuclear Nf κb Activity, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cardiomyocyte nuclear nf κb activity/product/Cayman Chemical
    Average 78 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cardiomyocyte nuclear nf κb activity - by Bioz Stars, 2020-01
    78/100 stars
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    79
    Cayman Chemical cardiomyocyte lysates
    <t>Cardiomyocyte</t> Isolation and General Plating Conditions.
    Cardiomyocyte Lysates, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 79/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cardiomyocyte lysates/product/Cayman Chemical
    Average 79 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    cardiomyocyte lysates - by Bioz Stars, 2020-01
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    Image Search Results


    Effect of doxorubicin and DoxQ on adult rat cardiomyocyte viability. a Dox versus DoxQ dose response. N = 3; mean ± SEM. * P

    Journal: Drug delivery and translational research

    Article Title: Mechanistically elucidating the in vitro safety and efficacy of a novel doxorubicin derivative

    doi: 10.1007/s13346-017-0379-2

    Figure Lengend Snippet: Effect of doxorubicin and DoxQ on adult rat cardiomyocyte viability. a Dox versus DoxQ dose response. N = 3; mean ± SEM. * P

    Article Snippet: The antioxidant activity of DoxQ, doxorubicin, quercetin, or a mixture of doxorubicin and quercetin was examined using the antioxidant kit from Cayman Chemical.

    Techniques:

    Fold change in P-gp ATPase activity of doxorubicin, DoxQ, a mixture of doxorubicin and quercetin, or quercetin at 1, 10, 50, 100, and 200 µM relative to basal activity. N = 4; mean ± SEM. * P

    Journal: Drug delivery and translational research

    Article Title: Mechanistically elucidating the in vitro safety and efficacy of a novel doxorubicin derivative

    doi: 10.1007/s13346-017-0379-2

    Figure Lengend Snippet: Fold change in P-gp ATPase activity of doxorubicin, DoxQ, a mixture of doxorubicin and quercetin, or quercetin at 1, 10, 50, 100, and 200 µM relative to basal activity. N = 4; mean ± SEM. * P

    Article Snippet: The antioxidant activity of DoxQ, doxorubicin, quercetin, or a mixture of doxorubicin and quercetin was examined using the antioxidant kit from Cayman Chemical.

    Techniques: Activity Assay

    The cytotoxic effects of doxorubicin, a mixture of doxorubicin and quercetin, or DoxQ determined by resazurin blue assay expressed as IC 50 after 5 µM treatment for 72 h. N = 4; mean ± SD

    Journal: Drug delivery and translational research

    Article Title: Mechanistically elucidating the in vitro safety and efficacy of a novel doxorubicin derivative

    doi: 10.1007/s13346-017-0379-2

    Figure Lengend Snippet: The cytotoxic effects of doxorubicin, a mixture of doxorubicin and quercetin, or DoxQ determined by resazurin blue assay expressed as IC 50 after 5 µM treatment for 72 h. N = 4; mean ± SD

    Article Snippet: The antioxidant activity of DoxQ, doxorubicin, quercetin, or a mixture of doxorubicin and quercetin was examined using the antioxidant kit from Cayman Chemical.

    Techniques:

    Fold expression of oxidative stress markers in RL-14 cells after treatment at 10 µM for 24 h of doxorubicin, DoxQ, or quercetin. The mRNA expression of a GST-A1 and b HO-1 in RL-14 cells. The mRNA expression was quantified by RT-PCR and normalized to β-actin as a housekeeping gene. N = 6; mean ± SEM. * P

    Journal: Drug delivery and translational research

    Article Title: Mechanistically elucidating the in vitro safety and efficacy of a novel doxorubicin derivative

    doi: 10.1007/s13346-017-0379-2

    Figure Lengend Snippet: Fold expression of oxidative stress markers in RL-14 cells after treatment at 10 µM for 24 h of doxorubicin, DoxQ, or quercetin. The mRNA expression of a GST-A1 and b HO-1 in RL-14 cells. The mRNA expression was quantified by RT-PCR and normalized to β-actin as a housekeeping gene. N = 6; mean ± SEM. * P

    Article Snippet: The antioxidant activity of DoxQ, doxorubicin, quercetin, or a mixture of doxorubicin and quercetin was examined using the antioxidant kit from Cayman Chemical.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    A schematic representation of the release of doxorubicin and quercetin from DoxQ

    Journal: Drug delivery and translational research

    Article Title: Mechanistically elucidating the in vitro safety and efficacy of a novel doxorubicin derivative

    doi: 10.1007/s13346-017-0379-2

    Figure Lengend Snippet: A schematic representation of the release of doxorubicin and quercetin from DoxQ

    Article Snippet: The antioxidant activity of DoxQ, doxorubicin, quercetin, or a mixture of doxorubicin and quercetin was examined using the antioxidant kit from Cayman Chemical.

    Techniques:

    In vitro release of doxorubicin and quercetin from DoxQ quantified by HPLC. N = 3; mean ± SEM

    Journal: Drug delivery and translational research

    Article Title: Mechanistically elucidating the in vitro safety and efficacy of a novel doxorubicin derivative

    doi: 10.1007/s13346-017-0379-2

    Figure Lengend Snippet: In vitro release of doxorubicin and quercetin from DoxQ quantified by HPLC. N = 3; mean ± SEM

    Article Snippet: The antioxidant activity of DoxQ, doxorubicin, quercetin, or a mixture of doxorubicin and quercetin was examined using the antioxidant kit from Cayman Chemical.

    Techniques: In Vitro, High Performance Liquid Chromatography

    Fluorescence imaging study of cell uptake of doxorubicin by MDCK-MDR cells (P-gp positive). Cells were treated with a 50 nM free doxorubicin, b 50 nM doxorubicin and 50 nM quercetin, and c 50 nM DoxQ

    Journal: Drug delivery and translational research

    Article Title: Mechanistically elucidating the in vitro safety and efficacy of a novel doxorubicin derivative

    doi: 10.1007/s13346-017-0379-2

    Figure Lengend Snippet: Fluorescence imaging study of cell uptake of doxorubicin by MDCK-MDR cells (P-gp positive). Cells were treated with a 50 nM free doxorubicin, b 50 nM doxorubicin and 50 nM quercetin, and c 50 nM DoxQ

    Article Snippet: The antioxidant activity of DoxQ, doxorubicin, quercetin, or a mixture of doxorubicin and quercetin was examined using the antioxidant kit from Cayman Chemical.

    Techniques: Fluorescence, Imaging

    Antioxidant activity of DoxQ in comparison to doxorubicin, a mixture of doxorubicin and quercetin, or quercetin alone expressed as Trolox equivalents ( N = 4; mean ± SEM). + P

    Journal: Drug delivery and translational research

    Article Title: Mechanistically elucidating the in vitro safety and efficacy of a novel doxorubicin derivative

    doi: 10.1007/s13346-017-0379-2

    Figure Lengend Snippet: Antioxidant activity of DoxQ in comparison to doxorubicin, a mixture of doxorubicin and quercetin, or quercetin alone expressed as Trolox equivalents ( N = 4; mean ± SEM). + P

    Article Snippet: The antioxidant activity of DoxQ, doxorubicin, quercetin, or a mixture of doxorubicin and quercetin was examined using the antioxidant kit from Cayman Chemical.

    Techniques: Antioxidant Activity Assay

    Fold expression of cardiac hypertrophy markers in RL-14 cells after treatment at 10 µM for 24 h of doxorubicin, DoxQ, or quercetin. The mRNA expression of a BNP, b α-MHC, and c β-MHC in RL-14 cells. The mRNA expression was quantified by RT-PCR and normalized to β-actin as a housekeeping gene. N = 6; mean ± SEM.* P

    Journal: Drug delivery and translational research

    Article Title: Mechanistically elucidating the in vitro safety and efficacy of a novel doxorubicin derivative

    doi: 10.1007/s13346-017-0379-2

    Figure Lengend Snippet: Fold expression of cardiac hypertrophy markers in RL-14 cells after treatment at 10 µM for 24 h of doxorubicin, DoxQ, or quercetin. The mRNA expression of a BNP, b α-MHC, and c β-MHC in RL-14 cells. The mRNA expression was quantified by RT-PCR and normalized to β-actin as a housekeeping gene. N = 6; mean ± SEM.* P

    Article Snippet: The antioxidant activity of DoxQ, doxorubicin, quercetin, or a mixture of doxorubicin and quercetin was examined using the antioxidant kit from Cayman Chemical.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    Effects of dobutamine (DOB) and lipopolysaccharide (LPS) on cardiomyocyte Bcl-2 levels, mitochondrial Bax contents, mitochondrial membrane potential and cytochrome c release. Adult mouse ventricular myocytes were treated with DOB (0.02 μM), LPS (10 ng/mL), their combination, or vehicle for 12 hours. (A) Bcl-2 levels were analyzed by western blotting of whole cell lysates. (B) Mitochondrial Bax protein contents were detected by western blotting of mitochondrial proteins and voltage-dependent anion channel (VDAC) was immunoblotted as a mitochondrial marker. (C) Confocal images of JC-1 fluorescence. Mitochondrial membrane potential was visualized in ventricular myocytes stained with JC-1, an indicator of mitochondrial function, green fluorescence indicates monomeric JC-1 (left), red fluorescence represents aggregate JC-1 (middle) and merged channels are shown (right): scale bar = 20 μm. (D) The ratio of monomeric and aggregated JC-1, indicating changes in mitochondrial membrane potential. (E) Cytosolic cytochrome c (Cyt c) contents were analyzed by western blotting of cytosolic proteins and GAPDH was used as an internal control: mean ± standard error of the mean (n = 3). * P

    Journal: Critical Care

    Article Title: β1-adrenoceptor stimulation promotes LPS-induced cardiomyocyte apoptosis through activating PKA and enhancing CaMKII and IκBα phosphorylation

    doi: 10.1186/s13054-015-0820-1

    Figure Lengend Snippet: Effects of dobutamine (DOB) and lipopolysaccharide (LPS) on cardiomyocyte Bcl-2 levels, mitochondrial Bax contents, mitochondrial membrane potential and cytochrome c release. Adult mouse ventricular myocytes were treated with DOB (0.02 μM), LPS (10 ng/mL), their combination, or vehicle for 12 hours. (A) Bcl-2 levels were analyzed by western blotting of whole cell lysates. (B) Mitochondrial Bax protein contents were detected by western blotting of mitochondrial proteins and voltage-dependent anion channel (VDAC) was immunoblotted as a mitochondrial marker. (C) Confocal images of JC-1 fluorescence. Mitochondrial membrane potential was visualized in ventricular myocytes stained with JC-1, an indicator of mitochondrial function, green fluorescence indicates monomeric JC-1 (left), red fluorescence represents aggregate JC-1 (middle) and merged channels are shown (right): scale bar = 20 μm. (D) The ratio of monomeric and aggregated JC-1, indicating changes in mitochondrial membrane potential. (E) Cytosolic cytochrome c (Cyt c) contents were analyzed by western blotting of cytosolic proteins and GAPDH was used as an internal control: mean ± standard error of the mean (n = 3). * P

    Article Snippet: Cardiomyocyte nuclear NF-κB activity was measured using a NF-kB (p65) Transcription Factor Assay kit (Cayman Chemical, Ann Arbor, MI, USA) as previously described [ ].

    Techniques: Western Blot, Marker, Fluorescence, Staining

    Dobutamine (DOB) enhances caspase-8 and caspase-9 activation in lipopolysaccharide (LPS)-treated adult mouse cardiomyocytes. Caspase-8 (A) and caspase-9 (B) activities were detected in cardiomyocytes treated with 0.02 μM DOB or/and 10 ng/mL LPS for 6, 12 or 24 hours. Mean ± standard error of the mean (n = 4). * P

    Journal: Critical Care

    Article Title: β1-adrenoceptor stimulation promotes LPS-induced cardiomyocyte apoptosis through activating PKA and enhancing CaMKII and IκBα phosphorylation

    doi: 10.1186/s13054-015-0820-1

    Figure Lengend Snippet: Dobutamine (DOB) enhances caspase-8 and caspase-9 activation in lipopolysaccharide (LPS)-treated adult mouse cardiomyocytes. Caspase-8 (A) and caspase-9 (B) activities were detected in cardiomyocytes treated with 0.02 μM DOB or/and 10 ng/mL LPS for 6, 12 or 24 hours. Mean ± standard error of the mean (n = 4). * P

    Article Snippet: Cardiomyocyte nuclear NF-κB activity was measured using a NF-kB (p65) Transcription Factor Assay kit (Cayman Chemical, Ann Arbor, MI, USA) as previously described [ ].

    Techniques: Activation Assay

    Proposed signaling mechanisms for lipopolysaccharide (LPS)-induced cardiomyocyte apoptosis enhanced by β 1 -AR activation. Cardiomyocyte β 1 -AR activation augments IκBα phosphorylation and TNF-α expression, activates protein kinase A (PKA), enhances calmodulin-dependent protein kinase II (CaMKII) phosphorylation, and subsequently promotes LPS-induced cardiomyocyte apoptosis. Cardiomyocyte PKA activation enhances p38 mitogen-activated protein kinase (MAPK) and c-jun NH2-terminal kinase (JNK) phosphorylation, reduces Bcl-2 protein levels, and in turn leads to an increase in cytochrome c release and caspase-9 activity during LPS challenge.

    Journal: Critical Care

    Article Title: β1-adrenoceptor stimulation promotes LPS-induced cardiomyocyte apoptosis through activating PKA and enhancing CaMKII and IκBα phosphorylation

    doi: 10.1186/s13054-015-0820-1

    Figure Lengend Snippet: Proposed signaling mechanisms for lipopolysaccharide (LPS)-induced cardiomyocyte apoptosis enhanced by β 1 -AR activation. Cardiomyocyte β 1 -AR activation augments IκBα phosphorylation and TNF-α expression, activates protein kinase A (PKA), enhances calmodulin-dependent protein kinase II (CaMKII) phosphorylation, and subsequently promotes LPS-induced cardiomyocyte apoptosis. Cardiomyocyte PKA activation enhances p38 mitogen-activated protein kinase (MAPK) and c-jun NH2-terminal kinase (JNK) phosphorylation, reduces Bcl-2 protein levels, and in turn leads to an increase in cytochrome c release and caspase-9 activity during LPS challenge.

    Article Snippet: Cardiomyocyte nuclear NF-κB activity was measured using a NF-kB (p65) Transcription Factor Assay kit (Cayman Chemical, Ann Arbor, MI, USA) as previously described [ ].

    Techniques: Activation Assay, Expressing, Activity Assay

    Atenolol (ATE) inhibits cardiomyocyte apoptosis in lipolysaccharide (LPS)-challenged mice. Animals received intraperitoneal injection of normal saline or ATE (10 mg/kg) 1 hour before normal saline or 20 mg/kg LPS administration. At 12 hours after LPS administration, cardiomyocyte apoptosis was examined by TUNEL assay. (A) Representative photomicrographs of TUNEL assay (green) in the hearts from LPS and ATE plus LPS groups. Total nuclei were stained with DAPI (blue) and cardiomyocytes with anti-cTnI antibody (red). (B) Apoptotic index (AI) of cardiomyocytes (mean ± standard error of the mean (SEM); n = 8). (C) Cardiac caspase 3/7 activity at 6 hours after LPS administration (mean ± SEM; n = 8). * P

    Journal: Critical Care

    Article Title: β1-adrenoceptor stimulation promotes LPS-induced cardiomyocyte apoptosis through activating PKA and enhancing CaMKII and IκBα phosphorylation

    doi: 10.1186/s13054-015-0820-1

    Figure Lengend Snippet: Atenolol (ATE) inhibits cardiomyocyte apoptosis in lipolysaccharide (LPS)-challenged mice. Animals received intraperitoneal injection of normal saline or ATE (10 mg/kg) 1 hour before normal saline or 20 mg/kg LPS administration. At 12 hours after LPS administration, cardiomyocyte apoptosis was examined by TUNEL assay. (A) Representative photomicrographs of TUNEL assay (green) in the hearts from LPS and ATE plus LPS groups. Total nuclei were stained with DAPI (blue) and cardiomyocytes with anti-cTnI antibody (red). (B) Apoptotic index (AI) of cardiomyocytes (mean ± standard error of the mean (SEM); n = 8). (C) Cardiac caspase 3/7 activity at 6 hours after LPS administration (mean ± SEM; n = 8). * P

    Article Snippet: Cardiomyocyte nuclear NF-κB activity was measured using a NF-kB (p65) Transcription Factor Assay kit (Cayman Chemical, Ann Arbor, MI, USA) as previously described [ ].

    Techniques: Mouse Assay, Injection, TUNEL Assay, Staining, Activity Assay

    Dobutamine promotes TNF-α expression and phosphorylation of IκBα, p38 mitogen-activated protein kinase (MAPK) and c-jun NH2-terminal kinase (JNK) in lipopolysaccharide (LPS)-treated cardiomyocytes. Adult mouse ventricular myocytes were treated with dobutamine (DOB, 0.02 μM), LPS (10 ng/mL), their combination, or vehicle for 6 hours and 12 hours. The total cardiomyocyte lysates and nuclear proteins were prepared. (A) The expression of TNF-α in cardiomyocytes was detected by western blot (mean ± standard error of the mean (SEM); n = 6). (B) NF-kB p65 DNA binding activity in nuclear proteins was examined using Cayman’s NF-kB (p65) Transcription Factor Assay kit, and is expressed as percent changes relative to control (mean ± SEM; n = 6). The levels of p-IκBα/IκBα (C) , p-ERK/ERK (D) , p-p38 MAPK/ p38 MAPK (E) and p-JNK/JNK (F) were examined by western blotting of whole cell lysates with specific antibodies, respectively (mean ± SEM; n = 3). * P

    Journal: Critical Care

    Article Title: β1-adrenoceptor stimulation promotes LPS-induced cardiomyocyte apoptosis through activating PKA and enhancing CaMKII and IκBα phosphorylation

    doi: 10.1186/s13054-015-0820-1

    Figure Lengend Snippet: Dobutamine promotes TNF-α expression and phosphorylation of IκBα, p38 mitogen-activated protein kinase (MAPK) and c-jun NH2-terminal kinase (JNK) in lipopolysaccharide (LPS)-treated cardiomyocytes. Adult mouse ventricular myocytes were treated with dobutamine (DOB, 0.02 μM), LPS (10 ng/mL), their combination, or vehicle for 6 hours and 12 hours. The total cardiomyocyte lysates and nuclear proteins were prepared. (A) The expression of TNF-α in cardiomyocytes was detected by western blot (mean ± standard error of the mean (SEM); n = 6). (B) NF-kB p65 DNA binding activity in nuclear proteins was examined using Cayman’s NF-kB (p65) Transcription Factor Assay kit, and is expressed as percent changes relative to control (mean ± SEM; n = 6). The levels of p-IκBα/IκBα (C) , p-ERK/ERK (D) , p-p38 MAPK/ p38 MAPK (E) and p-JNK/JNK (F) were examined by western blotting of whole cell lysates with specific antibodies, respectively (mean ± SEM; n = 3). * P

    Article Snippet: Cardiomyocyte nuclear NF-κB activity was measured using a NF-kB (p65) Transcription Factor Assay kit (Cayman Chemical, Ann Arbor, MI, USA) as previously described [ ].

    Techniques: Expressing, Western Blot, Binding Assay, Activity Assay, Transcription Factor Assay

    Effects of KT5720, a protein kinase A (PKA) inhibitor, on apoptosis-associated molecules in dobutamine (DOB) and lipolysaccharide (LPS)-treated cardiomyocytes. Adult mouse ventricular myocytes were pretreated with KT5720 (5 μM) or vehicle for 1 hour, and then exposed to DOB (0.02 μM), LPS (10 ng/mL), their combination, or vehicle for 12 hours. Caspase-8 (A) and caspase-9 (C) activities were detected 12 hours after DOB and LPS treatment (mean ± standard error of the mean (SEM); n = 8). IκBα (B), p38 mitogen-activated protein kinase (MAPK) (D) and c-jun NH2-terminal kinase (JNK) (E) phosphorylation, as well as Bcl-2 (F) protein expression in cardiomyocytes, were also examined 12 hours after DOB and LPS treatment using western blotting (mean ± SEM; n = 3). The results showed that KT5720 reversed the effects of dobutamine (DOB) on caspase-9 activation, Bcl-2 expression as well as p38 MAPK and JNK phosphorylation, but not on caspase-8 activation and IκBα phosphorylation in LPS-challenged cardiomyocytes. * P

    Journal: Critical Care

    Article Title: β1-adrenoceptor stimulation promotes LPS-induced cardiomyocyte apoptosis through activating PKA and enhancing CaMKII and IκBα phosphorylation

    doi: 10.1186/s13054-015-0820-1

    Figure Lengend Snippet: Effects of KT5720, a protein kinase A (PKA) inhibitor, on apoptosis-associated molecules in dobutamine (DOB) and lipolysaccharide (LPS)-treated cardiomyocytes. Adult mouse ventricular myocytes were pretreated with KT5720 (5 μM) or vehicle for 1 hour, and then exposed to DOB (0.02 μM), LPS (10 ng/mL), their combination, or vehicle for 12 hours. Caspase-8 (A) and caspase-9 (C) activities were detected 12 hours after DOB and LPS treatment (mean ± standard error of the mean (SEM); n = 8). IκBα (B), p38 mitogen-activated protein kinase (MAPK) (D) and c-jun NH2-terminal kinase (JNK) (E) phosphorylation, as well as Bcl-2 (F) protein expression in cardiomyocytes, were also examined 12 hours after DOB and LPS treatment using western blotting (mean ± SEM; n = 3). The results showed that KT5720 reversed the effects of dobutamine (DOB) on caspase-9 activation, Bcl-2 expression as well as p38 MAPK and JNK phosphorylation, but not on caspase-8 activation and IκBα phosphorylation in LPS-challenged cardiomyocytes. * P

    Article Snippet: Cardiomyocyte nuclear NF-κB activity was measured using a NF-kB (p65) Transcription Factor Assay kit (Cayman Chemical, Ann Arbor, MI, USA) as previously described [ ].

    Techniques: Expressing, Western Blot, Activation Assay

    Caspase 3/7 activity and apoptotic index of cardiomyocytes treated with dobutamine (DOB) and lipopolysaccharide (LPS). (A,B) Adult mouse ventricular myocytes treated with DOB at various doses for 24 hours. (C,D) Adult mouse ventricular myocytes exposed to LPS at various concentrations for 24 hours. (E) Representative confocal images of TUNEL assay (green) of adult mouse ventricular myocytes treated with 0.02 μM DOB or/and 10 ng/ml LPS for 24 hours; all cardiomyocytes were stained with anti-cardiac troponin I (cTnI) antibody (red) and total nuclei with 4', 6-diamidino-2-phenylindole (DAPI, blue). White arrowheads indicate apoptotic cardiomyocytes (scale bar = 20 μm). (F,G) AI and caspase-3/7 activity in cardiomyocytes treated with 0.02 μM DOB or/and 10 ng/mL LPS for 24 hours. Mean ± standard error of the mean (n = 4 to 6). * P

    Journal: Critical Care

    Article Title: β1-adrenoceptor stimulation promotes LPS-induced cardiomyocyte apoptosis through activating PKA and enhancing CaMKII and IκBα phosphorylation

    doi: 10.1186/s13054-015-0820-1

    Figure Lengend Snippet: Caspase 3/7 activity and apoptotic index of cardiomyocytes treated with dobutamine (DOB) and lipopolysaccharide (LPS). (A,B) Adult mouse ventricular myocytes treated with DOB at various doses for 24 hours. (C,D) Adult mouse ventricular myocytes exposed to LPS at various concentrations for 24 hours. (E) Representative confocal images of TUNEL assay (green) of adult mouse ventricular myocytes treated with 0.02 μM DOB or/and 10 ng/ml LPS for 24 hours; all cardiomyocytes were stained with anti-cardiac troponin I (cTnI) antibody (red) and total nuclei with 4', 6-diamidino-2-phenylindole (DAPI, blue). White arrowheads indicate apoptotic cardiomyocytes (scale bar = 20 μm). (F,G) AI and caspase-3/7 activity in cardiomyocytes treated with 0.02 μM DOB or/and 10 ng/mL LPS for 24 hours. Mean ± standard error of the mean (n = 4 to 6). * P

    Article Snippet: Cardiomyocyte nuclear NF-κB activity was measured using a NF-kB (p65) Transcription Factor Assay kit (Cayman Chemical, Ann Arbor, MI, USA) as previously described [ ].

    Techniques: Activity Assay, TUNEL Assay, Staining

    Effects of dobutamine (DOB), lipolysachharide (LPS) and nifedipine (NIF) on cytosolic Ca 2+ , calmodulin-dependent protein kinase II (CaMKII) and caspase activation in cardiomyocytes. Adult mouse ventricular myocytes were pretreated with NIF (1 μM) , a calcium channel blocker, or vehicle for 1 hour, and then exposed to DOB (0.02 μM), LPS (10 ng/mL), their combination or vehicle for 3, 12 or 24 hours. (A,B) DOB enhanced LPS-induced cytosolic Ca 2+ concentration elevation (mean ± standard error of the mean (SEM); n = 8) and calmodulin-dependent protein kinase II (CaMKII) phosphorylation (mean ± SEM; n = 4) 3 hours and 12 hours after LPS treatment, respectively, both of which were blocked by NIF pretreatment. (C,D) NIF partly abolished the enhancement effects of DOB on LPS-stimulated caspase-9 (12 hours after DOB and LPS treatment) and caspase 3/7 (24 hours after DOB and LPS treatment) activation in cardiomyocytes (mean ± SEM; n = 8). * P

    Journal: Critical Care

    Article Title: β1-adrenoceptor stimulation promotes LPS-induced cardiomyocyte apoptosis through activating PKA and enhancing CaMKII and IκBα phosphorylation

    doi: 10.1186/s13054-015-0820-1

    Figure Lengend Snippet: Effects of dobutamine (DOB), lipolysachharide (LPS) and nifedipine (NIF) on cytosolic Ca 2+ , calmodulin-dependent protein kinase II (CaMKII) and caspase activation in cardiomyocytes. Adult mouse ventricular myocytes were pretreated with NIF (1 μM) , a calcium channel blocker, or vehicle for 1 hour, and then exposed to DOB (0.02 μM), LPS (10 ng/mL), their combination or vehicle for 3, 12 or 24 hours. (A,B) DOB enhanced LPS-induced cytosolic Ca 2+ concentration elevation (mean ± standard error of the mean (SEM); n = 8) and calmodulin-dependent protein kinase II (CaMKII) phosphorylation (mean ± SEM; n = 4) 3 hours and 12 hours after LPS treatment, respectively, both of which were blocked by NIF pretreatment. (C,D) NIF partly abolished the enhancement effects of DOB on LPS-stimulated caspase-9 (12 hours after DOB and LPS treatment) and caspase 3/7 (24 hours after DOB and LPS treatment) activation in cardiomyocytes (mean ± SEM; n = 8). * P

    Article Snippet: Cardiomyocyte nuclear NF-κB activity was measured using a NF-kB (p65) Transcription Factor Assay kit (Cayman Chemical, Ann Arbor, MI, USA) as previously described [ ].

    Techniques: Activation Assay, Concentration Assay

    Cardiomyocyte Isolation and General Plating Conditions.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Dystrophin–glycoprotein complex regulates muscle nitric oxide production through mechanoregulation of AMPK signaling

    doi: 10.1073/pnas.1512991112

    Figure Lengend Snippet: Cardiomyocyte Isolation and General Plating Conditions.

    Article Snippet: Stretched cardiomyocytes and nonstretched controls were then washed in DPBS and lysed in 0.1 M HCl for 20 min. Lysates were centrifuged at 1,000 × g for 10 min, and the supernatants were stored at −80 °C until use. cGMP concentration in cardiomyocyte lysates was analyzed using a cGMP EIA kit (Cayman Chemical; 581021) according to the manufacturer’s instructions, and was normalized to protein concentration based on the DC Protein Assay (Bio-Rad; 500-0116).

    Techniques: Isolation

    In Vitro Cardiomyocyte NO Imaging.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Dystrophin–glycoprotein complex regulates muscle nitric oxide production through mechanoregulation of AMPK signaling

    doi: 10.1073/pnas.1512991112

    Figure Lengend Snippet: In Vitro Cardiomyocyte NO Imaging.

    Article Snippet: Stretched cardiomyocytes and nonstretched controls were then washed in DPBS and lysed in 0.1 M HCl for 20 min. Lysates were centrifuged at 1,000 × g for 10 min, and the supernatants were stored at −80 °C until use. cGMP concentration in cardiomyocyte lysates was analyzed using a cGMP EIA kit (Cayman Chemical; 581021) according to the manufacturer’s instructions, and was normalized to protein concentration based on the DC Protein Assay (Bio-Rad; 500-0116).

    Techniques: In Vitro, Imaging

    nNOS localization. WT and mdx cardiomyocytes exhibit cytoplasmic and nuclear localization of nNOS (green) at rest. nNOS is not specifically enriched at the sarcolemma and does not colocalize with dystrophin (red). Cyclic stretch (1-Hz) does not lead to

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Dystrophin–glycoprotein complex regulates muscle nitric oxide production through mechanoregulation of AMPK signaling

    doi: 10.1073/pnas.1512991112

    Figure Lengend Snippet: nNOS localization. WT and mdx cardiomyocytes exhibit cytoplasmic and nuclear localization of nNOS (green) at rest. nNOS is not specifically enriched at the sarcolemma and does not colocalize with dystrophin (red). Cyclic stretch (1-Hz) does not lead to

    Article Snippet: Stretched cardiomyocytes and nonstretched controls were then washed in DPBS and lysed in 0.1 M HCl for 20 min. Lysates were centrifuged at 1,000 × g for 10 min, and the supernatants were stored at −80 °C until use. cGMP concentration in cardiomyocyte lysates was analyzed using a cGMP EIA kit (Cayman Chemical; 581021) according to the manufacturer’s instructions, and was normalized to protein concentration based on the DC Protein Assay (Bio-Rad; 500-0116).

    Techniques:

    AMPKα localization. WT and mdx cardiomyocytes exhibit cytoplasmic and nuclear localization of AMPKα (red) at rest. AMPKα is not specifically enriched at the sarcolemma and does not colocalize with dystrophin (green). Cyclic stretch

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Dystrophin–glycoprotein complex regulates muscle nitric oxide production through mechanoregulation of AMPK signaling

    doi: 10.1073/pnas.1512991112

    Figure Lengend Snippet: AMPKα localization. WT and mdx cardiomyocytes exhibit cytoplasmic and nuclear localization of AMPKα (red) at rest. AMPKα is not specifically enriched at the sarcolemma and does not colocalize with dystrophin (green). Cyclic stretch

    Article Snippet: Stretched cardiomyocytes and nonstretched controls were then washed in DPBS and lysed in 0.1 M HCl for 20 min. Lysates were centrifuged at 1,000 × g for 10 min, and the supernatants were stored at −80 °C until use. cGMP concentration in cardiomyocyte lysates was analyzed using a cGMP EIA kit (Cayman Chemical; 581021) according to the manufacturer’s instructions, and was normalized to protein concentration based on the DC Protein Assay (Bio-Rad; 500-0116).

    Techniques:

    Cardiomyocyte Lysis for Western Blot Analysis.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Dystrophin–glycoprotein complex regulates muscle nitric oxide production through mechanoregulation of AMPK signaling

    doi: 10.1073/pnas.1512991112

    Figure Lengend Snippet: Cardiomyocyte Lysis for Western Blot Analysis.

    Article Snippet: Stretched cardiomyocytes and nonstretched controls were then washed in DPBS and lysed in 0.1 M HCl for 20 min. Lysates were centrifuged at 1,000 × g for 10 min, and the supernatants were stored at −80 °C until use. cGMP concentration in cardiomyocyte lysates was analyzed using a cGMP EIA kit (Cayman Chemical; 581021) according to the manufacturer’s instructions, and was normalized to protein concentration based on the DC Protein Assay (Bio-Rad; 500-0116).

    Techniques: Lysis, Western Blot

    Stretch-dependent NO signaling is impaired in dystrophin-deficient cardiomyocytes. ( A ) Experimental setup. Adult mouse cardiomyocytes loaded with DAF-FM are visualized under 10x magnification using an epifluorescence microscope throughout 1 h of 1-Hz

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Dystrophin–glycoprotein complex regulates muscle nitric oxide production through mechanoregulation of AMPK signaling

    doi: 10.1073/pnas.1512991112

    Figure Lengend Snippet: Stretch-dependent NO signaling is impaired in dystrophin-deficient cardiomyocytes. ( A ) Experimental setup. Adult mouse cardiomyocytes loaded with DAF-FM are visualized under 10x magnification using an epifluorescence microscope throughout 1 h of 1-Hz

    Article Snippet: Stretched cardiomyocytes and nonstretched controls were then washed in DPBS and lysed in 0.1 M HCl for 20 min. Lysates were centrifuged at 1,000 × g for 10 min, and the supernatants were stored at −80 °C until use. cGMP concentration in cardiomyocyte lysates was analyzed using a cGMP EIA kit (Cayman Chemical; 581021) according to the manufacturer’s instructions, and was normalized to protein concentration based on the DC Protein Assay (Bio-Rad; 500-0116).

    Techniques: Microscopy

    Pharmacologic AMPK activation increases nNOS activity in dystrophin-deficient cardiomyocytes. ( A ) Treatment with AICAR elicits a significant increase in DAF-FM fluorescence in both WT and mdx cardiomyocytes (* P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Dystrophin–glycoprotein complex regulates muscle nitric oxide production through mechanoregulation of AMPK signaling

    doi: 10.1073/pnas.1512991112

    Figure Lengend Snippet: Pharmacologic AMPK activation increases nNOS activity in dystrophin-deficient cardiomyocytes. ( A ) Treatment with AICAR elicits a significant increase in DAF-FM fluorescence in both WT and mdx cardiomyocytes (* P

    Article Snippet: Stretched cardiomyocytes and nonstretched controls were then washed in DPBS and lysed in 0.1 M HCl for 20 min. Lysates were centrifuged at 1,000 × g for 10 min, and the supernatants were stored at −80 °C until use. cGMP concentration in cardiomyocyte lysates was analyzed using a cGMP EIA kit (Cayman Chemical; 581021) according to the manufacturer’s instructions, and was normalized to protein concentration based on the DC Protein Assay (Bio-Rad; 500-0116).

    Techniques: Activation Assay, Activity Assay, Fluorescence

    Pharmacologic AMPK activation increases stimulatory nNOS phosphorylation in dystrophin-deficient cardiomyocytes. ( A ) Acute treatment of WT and mdx cardiomyocytes with the AMPK activator AICAR increases activating phosphorylation of AMPKα at threonine

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Dystrophin–glycoprotein complex regulates muscle nitric oxide production through mechanoregulation of AMPK signaling

    doi: 10.1073/pnas.1512991112

    Figure Lengend Snippet: Pharmacologic AMPK activation increases stimulatory nNOS phosphorylation in dystrophin-deficient cardiomyocytes. ( A ) Acute treatment of WT and mdx cardiomyocytes with the AMPK activator AICAR increases activating phosphorylation of AMPKα at threonine

    Article Snippet: Stretched cardiomyocytes and nonstretched controls were then washed in DPBS and lysed in 0.1 M HCl for 20 min. Lysates were centrifuged at 1,000 × g for 10 min, and the supernatants were stored at −80 °C until use. cGMP concentration in cardiomyocyte lysates was analyzed using a cGMP EIA kit (Cayman Chemical; 581021) according to the manufacturer’s instructions, and was normalized to protein concentration based on the DC Protein Assay (Bio-Rad; 500-0116).

    Techniques: Activation Assay

    Stretch-induced nNOS and AMPK activation are impaired in dystrophin-deficient cardiomyocytes. ( A ) Acute stretch of WT cardiomyocytes elicits increased phosphorylation of nNOS at the stimulatory residue serine 1412, increased activating phosphorylation

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Dystrophin–glycoprotein complex regulates muscle nitric oxide production through mechanoregulation of AMPK signaling

    doi: 10.1073/pnas.1512991112

    Figure Lengend Snippet: Stretch-induced nNOS and AMPK activation are impaired in dystrophin-deficient cardiomyocytes. ( A ) Acute stretch of WT cardiomyocytes elicits increased phosphorylation of nNOS at the stimulatory residue serine 1412, increased activating phosphorylation

    Article Snippet: Stretched cardiomyocytes and nonstretched controls were then washed in DPBS and lysed in 0.1 M HCl for 20 min. Lysates were centrifuged at 1,000 × g for 10 min, and the supernatants were stored at −80 °C until use. cGMP concentration in cardiomyocyte lysates was analyzed using a cGMP EIA kit (Cayman Chemical; 581021) according to the manufacturer’s instructions, and was normalized to protein concentration based on the DC Protein Assay (Bio-Rad; 500-0116).

    Techniques: Activation Assay