fetal bovine serum fbs  (Thermo Fisher)


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    Structured Review

    Thermo Fisher fetal bovine serum fbs
    (A) Schematic diagram for the time line of CSCs induction. IM: Inducing medium; SM: Sphere culture medium. (B) Tumorsphere formation of different cancer cell lines. (a, b) Panc1 cells were induced with (a) or without (b) the small molecular compounds as described. In the presence of the compounds, cells formed typical tumorspheres (a); in contrast, in the absence of the compounds, only cell patches were observed (b). (c,d,e) Similarly, Bxpc3 (c), <t>H446</t> (d), and Ecs9706 (e) cells also formed spheres in suspension culture after 4 days induction. Scale bars, 100 μm. (C) Differentiation of induced tumorsphere cells. Induced panc1 sphere cells were cultured in suspension for 5 days. Cells were then digested into single cells and cultured in DMEM containing 10% <t>FBS</t> for 3 days. These cells restored their original morphology (upper panel) as parental panc1 cells (lower panel).
    Fetal Bovine Serum Fbs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 11498 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 11498 article reviews
    Price from $9.99 to $1999.99
    fetal bovine serum fbs - by Bioz Stars, 2020-02
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    Images

    1) Product Images from "Rapid induction of pancreatic cancer cells to cancer stem cells via heterochromatin modulation"

    Article Title: Rapid induction of pancreatic cancer cells to cancer stem cells via heterochromatin modulation

    Journal: Cell Cycle

    doi: 10.1080/15384101.2018.1489180

    (A) Schematic diagram for the time line of CSCs induction. IM: Inducing medium; SM: Sphere culture medium. (B) Tumorsphere formation of different cancer cell lines. (a, b) Panc1 cells were induced with (a) or without (b) the small molecular compounds as described. In the presence of the compounds, cells formed typical tumorspheres (a); in contrast, in the absence of the compounds, only cell patches were observed (b). (c,d,e) Similarly, Bxpc3 (c), H446 (d), and Ecs9706 (e) cells also formed spheres in suspension culture after 4 days induction. Scale bars, 100 μm. (C) Differentiation of induced tumorsphere cells. Induced panc1 sphere cells were cultured in suspension for 5 days. Cells were then digested into single cells and cultured in DMEM containing 10% FBS for 3 days. These cells restored their original morphology (upper panel) as parental panc1 cells (lower panel).
    Figure Legend Snippet: (A) Schematic diagram for the time line of CSCs induction. IM: Inducing medium; SM: Sphere culture medium. (B) Tumorsphere formation of different cancer cell lines. (a, b) Panc1 cells were induced with (a) or without (b) the small molecular compounds as described. In the presence of the compounds, cells formed typical tumorspheres (a); in contrast, in the absence of the compounds, only cell patches were observed (b). (c,d,e) Similarly, Bxpc3 (c), H446 (d), and Ecs9706 (e) cells also formed spheres in suspension culture after 4 days induction. Scale bars, 100 μm. (C) Differentiation of induced tumorsphere cells. Induced panc1 sphere cells were cultured in suspension for 5 days. Cells were then digested into single cells and cultured in DMEM containing 10% FBS for 3 days. These cells restored their original morphology (upper panel) as parental panc1 cells (lower panel).

    Techniques Used: Cell Culture

    Induction of heterochromatin alteration. Panc1 cells were cultured in inducing medium (induction) or DMEM containing 10% FBS (control) for 48 hrs. Immunofluorescence images show a significant reduction of heterochromatin markers H3K9me3 and HP1 in induced cells. DNA was counterstained by DAPI. Scale bar is equal to 10 μm.
    Figure Legend Snippet: Induction of heterochromatin alteration. Panc1 cells were cultured in inducing medium (induction) or DMEM containing 10% FBS (control) for 48 hrs. Immunofluorescence images show a significant reduction of heterochromatin markers H3K9me3 and HP1 in induced cells. DNA was counterstained by DAPI. Scale bar is equal to 10 μm.

    Techniques Used: Cell Culture, Immunofluorescence

    2) Product Images from "De novo lipogenesis represents a therapeutic target in mutant Kras non-small cell lung cancer"

    Article Title: De novo lipogenesis represents a therapeutic target in mutant Kras non-small cell lung cancer

    Journal: The FASEB Journal

    doi: 10.1096/fj.201800204

    Cells expressing mutant Kras are sensitized to FASN inhibitor-based growth inhibition. A , B ) Nontarget shRNA and Kras expressing shRNA A549 ( A ) and H23 ( B ) cells were treated with the indicated doses of C75 or GSK2194069, and the cell numbers were determined. C , D ) H838 ( C ) and H1299 ( D ) cells expressing GFP or Kras G12V were treated with the indicated doses of C75 or GSK2194069, and the cell numbers were determined using a Countess automated cell counter; n = 3–5. Experiments were performed in RPMI-1640 medium supplemented with 1% FBS. Shown is the percentage of cells after treatment with the indicated drugs compared with vehicle. * P
    Figure Legend Snippet: Cells expressing mutant Kras are sensitized to FASN inhibitor-based growth inhibition. A , B ) Nontarget shRNA and Kras expressing shRNA A549 ( A ) and H23 ( B ) cells were treated with the indicated doses of C75 or GSK2194069, and the cell numbers were determined. C , D ) H838 ( C ) and H1299 ( D ) cells expressing GFP or Kras G12V were treated with the indicated doses of C75 or GSK2194069, and the cell numbers were determined using a Countess automated cell counter; n = 3–5. Experiments were performed in RPMI-1640 medium supplemented with 1% FBS. Shown is the percentage of cells after treatment with the indicated drugs compared with vehicle. * P

    Techniques Used: Expressing, Mutagenesis, Inhibition, shRNA

    Palmitate rescues FASN inhibitor-mediated inhibition of cell growth. A549 ( A , C ) and H23 ( B , D ) expressing NTshRNA (left) or shKras (right) were treated with 2.5 μM GSK2194069 or 10 μM C75 alone or in combination with 7.5 μM palmitate. Cell number was determined with a Countess automated cell counter; n = 3. Experiments were performed in RPMI-1640 medium supplemented with 1% FBS. Bovine serum albumin was added as a lipid carrier because of the reduced serum conditions. * P
    Figure Legend Snippet: Palmitate rescues FASN inhibitor-mediated inhibition of cell growth. A549 ( A , C ) and H23 ( B , D ) expressing NTshRNA (left) or shKras (right) were treated with 2.5 μM GSK2194069 or 10 μM C75 alone or in combination with 7.5 μM palmitate. Cell number was determined with a Countess automated cell counter; n = 3. Experiments were performed in RPMI-1640 medium supplemented with 1% FBS. Bovine serum albumin was added as a lipid carrier because of the reduced serum conditions. * P

    Techniques Used: Inhibition, Expressing

    3) Product Images from "Preconditioning of mesenchymal stromal cells with low-intensity ultrasound: influence on chondrogenesis and directed SOX9 signaling pathways"

    Article Title: Preconditioning of mesenchymal stromal cells with low-intensity ultrasound: influence on chondrogenesis and directed SOX9 signaling pathways

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-019-1532-2

    The culture of MSC-laden HyStem-C constructs under cLIUS stimulation. MSCs were encapsulated at a density of 5 × 10 6 cells/ml of HyStem-C hydrogel and grown in DMEM medium supplemented with 10% FBS, 100 nM dexamethasone, and 50 μg/ml l -ascorbic acid for 6 weeks under cLIUS at 14 kPa (5 MHz, 2.5 Vpp), 20 min/application, and 4 applications/day ( n = 3). Non-cLIUS-stimulated 3D constructs served as controls ( n = 3). Representative images of 4-μm sections of the constructs stained immunohistochemically for collagen II and chondroitin sulfate is shown. Scale bar represents 100 μm
    Figure Legend Snippet: The culture of MSC-laden HyStem-C constructs under cLIUS stimulation. MSCs were encapsulated at a density of 5 × 10 6 cells/ml of HyStem-C hydrogel and grown in DMEM medium supplemented with 10% FBS, 100 nM dexamethasone, and 50 μg/ml l -ascorbic acid for 6 weeks under cLIUS at 14 kPa (5 MHz, 2.5 Vpp), 20 min/application, and 4 applications/day ( n = 3). Non-cLIUS-stimulated 3D constructs served as controls ( n = 3). Representative images of 4-μm sections of the constructs stained immunohistochemically for collagen II and chondroitin sulfate is shown. Scale bar represents 100 μm

    Techniques Used: Construct, Staining

    4) Product Images from "Anti-Oxidative and Anti-Inflammatory Activity of Kenya Grade AA Green Coffee Bean Extracts"

    Article Title: Anti-Oxidative and Anti-Inflammatory Activity of Kenya Grade AA Green Coffee Bean Extracts

    Journal: Iranian Journal of Public Health

    doi:

    Cell viability of Kenya AA green coffee bean extracts on macrophage cells (RAW 264.7) RAW 264.7 cells were incubated for 24 hours in DMEM containing 10 % FBS and treated with increasing concentrations of Kenya AA green coffee bean extracts for 24 hours and cell viability was measured by MTT assays. Each value represents mean ± standard deviation of three individual experiments. ** P
    Figure Legend Snippet: Cell viability of Kenya AA green coffee bean extracts on macrophage cells (RAW 264.7) RAW 264.7 cells were incubated for 24 hours in DMEM containing 10 % FBS and treated with increasing concentrations of Kenya AA green coffee bean extracts for 24 hours and cell viability was measured by MTT assays. Each value represents mean ± standard deviation of three individual experiments. ** P

    Techniques Used: Incubation, MTT Assay, Standard Deviation

    5) Product Images from "Anti-Oxidative and Anti-Inflammatory Activity of Kenya Grade AA Green Coffee Bean Extracts"

    Article Title: Anti-Oxidative and Anti-Inflammatory Activity of Kenya Grade AA Green Coffee Bean Extracts

    Journal: Iranian Journal of Public Health

    doi:

    Cell viability of Kenya AA green coffee bean extracts on macrophage cells (RAW 264.7) RAW 264.7 cells were incubated for 24 hours in DMEM containing 10 % FBS and treated with increasing concentrations of Kenya AA green coffee bean extracts for 24 hours and cell viability was measured by MTT assays. Each value represents mean ± standard deviation of three individual experiments. ** P
    Figure Legend Snippet: Cell viability of Kenya AA green coffee bean extracts on macrophage cells (RAW 264.7) RAW 264.7 cells were incubated for 24 hours in DMEM containing 10 % FBS and treated with increasing concentrations of Kenya AA green coffee bean extracts for 24 hours and cell viability was measured by MTT assays. Each value represents mean ± standard deviation of three individual experiments. ** P

    Techniques Used: Incubation, MTT Assay, Standard Deviation

    Related Articles

    Transduction:

    Article Title: De novo lipogenesis represents a therapeutic target in mutant Kras non-small cell lung cancer
    Article Snippet: A549 and H23 NSCLC cells were transduced with lentivirus expressing nontarget short hairpin RNA (shRNA) pLKO (A549NTshRNA and H23NTshRNA ) or shRNA against Kras (A549sh Kras and H23sh Kras ; TRCN0000033260,TRCN0000033262, TRCN0000033261, and TRCN0000033259; MilliporeSigma, Burlington, MA, USA). .. For cell proliferation, cells were seeded into 6- or 24-well plates overnight and then treated with the indicated doses of inhibitors using Roswell Park Memorial Institute (RPMI)-1640 (Buffalo, NY, USA) medium supplemented with either 1% fetal bovine serum (FBS), and the number of cells was determined after 3–5 d by hemocytometer or Countess Automated Cell Counter (Thermo Fisher Scientific, Waltham, MA, USA).

    Stable Transfection:

    Article Title: Protein kinase D1 regulates ERα-positive breast cancer cell growth response to 17β-estradiol and contributes to poor prognosis in patients
    Article Snippet: The MCF-7 cell line was cultured in DMEM-Glutamax®, supplemented with 10% foetal bovine serum (FBS) and 100 units/ml penicillin and 100 μg/ml streptomycin (P/S); complete medium (Invitrogen-Life Technologies, Cergy-Pontoise, France). .. 1 mg/ml G418 (Calbiochem, Darmstadt, Germany) was added to this medium for stably transfected MCF-7 cell culture.

    Construct:

    Article Title: Preconditioning of mesenchymal stromal cells with low-intensity ultrasound: influence on chondrogenesis and directed SOX9 signaling pathways
    Article Snippet: .. The inserts were then carefully removed after the solidification of MSC-laden hydrogels which typically ensued within (two constructs per well) were cultured in six-well tissue culture plates (TCP) in DMEM-high glucose medium supplemented with 10% fetal bovine serum (FBS) (Gibco, USA), 100 nM dexamethasone, 50 μg/ml ascorbic acid, and 1× antibiotic-antimycotic solution (Gibco, USA) at 37 °C and 5% CO2 with or without cLIUS for 6 weeks. .. cLIUS treatment regimen of 3D MSC constructs Six-well TCPs containing MSC-hydrogel constructs were placed in plate holders of a cLIUS-assisted incubator developed at the Department of Chemical Engineering, University of Nebraska-Lincoln (UNL), USA, with operating procedures detailed elsewhere [ ]. cLIUS was applied to the plates at 5 MHz frequency and 2.5 Vpp (14 kPa) for 20 min per application at four applications/day for a period of 6 weeks.

    Transplantation Assay:

    Article Title: Fibulin1C peptide induces cell attachment and extracellular matrix deposition in lung fibroblasts
    Article Snippet: Briefly, ASM cells were isolated from medium to large airways and fibroblasts were derived from proximal lung tissues which were obtained from macroscopically normal surgical tailings following resection for thoracic lesions, and from lung transplantation for emphysema or pulmonary fibrosis. .. ASM cells and fibroblasts were maintained in Dulbecco's Modified Eagle Medium (DMEM) (Gibco, Grand Island, New York, US) with 5% foetal bovine serum (FBS) and 1% Antibiotic-Antimycotic (Gibco, Grand Island, New York, US).

    Incubation:

    Article Title: Rapid induction of pancreatic cancer cells to cancer stem cells via heterochromatin modulation
    Article Snippet: Panc1 and H446 cells were cultured in DMEM (Gibco-life Technologies, USA) containing 10% fetal bovine serum (FBS) (Gibco/Invitrogen, Australia), 1% penicillin/streptomycin (PS) (Invitrogen). .. All the cells were incubated at 37℃ in a humidified incubator in the presence of 5% CO2.

    Article Title: New Proof-of-Concept in Viral Inactivation: Virucidal Efficacy of 405 nm Light Against Feline Calicivirus as a Model for Norovirus Decontamination
    Article Snippet: Cell and Virus Culture Feline embryonic cells, strain FEA (Jarrett et al. ), were cultured in Dulbecco’s modified eagle’s medium (DMEM) supplemented with 10% foetal bovine serum (FBS), 2 mM l -glutamine, 1 mM sodium pyruvate and 240 U mL−1 penicillin streptomycin (Gibco, Life Technologies, UK), to form 10% FBS-DMEM. .. After 90 min incubation of the inoculated cells on a rotating roller stand at 37 °C in 5% CO2 , fresh culture medium was added and flasks incubated for 24 h. This resulted in virus-induced destruction of nearly 90% of the cell monolayer.

    Article Title: Four Cysteine Residues Contribute to Homodimerization of Chicken Interleukin-2
    Article Snippet: .. Equal amounts of the chIL-2 dimer were incubated in PBS buffer containing dissociation reagents in a series of five-fold dilutions from the initial concentrations of 4 M KSCN [ ], 10% sodium cholate [ ], 20% Triton X-100 [ ], 8 M urea, 4.8 M guanidine hydrochloride [ ], 215 mM EDTA [ ], 6 M β-ME and 0.8 M DTT, at 37 °C for 1 h. To test the effect of foetal bovine serum (FBS) on dimerization of chIL-2, freshly purified chIL-2 (mostly monomeric) and chIL-2 dimer (post incubation at 37 °C for 24 h) were added to the T cell culture medium (RPMI 1640 medium supplemented with 10% FBS (Gibco, Waltham, MA, USA), 100 U penicillin, and 100 mg streptomycin (Gibco)) and incubated at 37 °C for 24 h. Western blot was employed for the detection of chIL-2. ..

    Expressing:

    Article Title: Protein kinase D1 regulates ERα-positive breast cancer cell growth response to 17β-estradiol and contributes to poor prognosis in patients
    Article Snippet: For PKD1 expression and clinicopathological analyses, each of the 38 cell lines tested was cultured in the conditions recommended by ATCC. .. The MCF-7 cell line was cultured in DMEM-Glutamax®, supplemented with 10% foetal bovine serum (FBS) and 100 units/ml penicillin and 100 μg/ml streptomycin (P/S); complete medium (Invitrogen-Life Technologies, Cergy-Pontoise, France).

    Article Title: De novo lipogenesis represents a therapeutic target in mutant Kras non-small cell lung cancer
    Article Snippet: A549 and H23 NSCLC cells were transduced with lentivirus expressing nontarget short hairpin RNA (shRNA) pLKO (A549NTshRNA and H23NTshRNA ) or shRNA against Kras (A549sh Kras and H23sh Kras ; TRCN0000033260,TRCN0000033262, TRCN0000033261, and TRCN0000033259; MilliporeSigma, Burlington, MA, USA). .. For cell proliferation, cells were seeded into 6- or 24-well plates overnight and then treated with the indicated doses of inhibitors using Roswell Park Memorial Institute (RPMI)-1640 (Buffalo, NY, USA) medium supplemented with either 1% fetal bovine serum (FBS), and the number of cells was determined after 3–5 d by hemocytometer or Countess Automated Cell Counter (Thermo Fisher Scientific, Waltham, MA, USA).

    Article Title: Anti-Oxidative and Anti-Inflammatory Activity of Kenya Grade AA Green Coffee Bean Extracts
    Article Snippet: Cell line, Raw 264.7, a macrophage that is used for cell culture and cell toxicity measurement was purchased from Korea cell line bank (Korea Cell Line Bank, Seoul, Korea), and Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), phosphate buffered saline (PBS), penicillin/streptomycin, trypsin for cell culture was purchased from Thermo Scientific HyClone (Logan, UT, USA). .. Furthermore, primary antibody iNOS, COX-2 and secondary antibody anti-mouse for protein expression measurement were purchased from Santa Cruz (CA, USA).

    Modification:

    Article Title: The interaction of fluorescent nanodiamond probes with cellular media
    Article Snippet: Aggregate/corona formation We chose to analyze the interaction with DMEM (Dulbecco’s Modified Eagle Medium) since it is the most common standard cell medium for culturing mammalian cells. .. The different media were (1) DMEM Complete (consisting of DMEM + complements: Glutamax (1%), Pen/Strep (1%), Foetal bovine serum (FBS) (10%), Gibco Life Technologies, Bleiswijk, the Netherlands, www.thermofisher.com/ch/en/home/brands/gibco ). (2) DMEM without complements and (3) pure or diluted (10%) FBS.

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways
    Article Snippet: .. Reagents Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA). .. Calcium chloride dehydrate (CaCl2 ·2H2 O), sodium bicarbonate, hepes, dimethyl sulphoxide (DMSO) and thiazolyl blue tetrazolium bromide (MTT) were from Sigma-Aldrich (St. Louis, MO, USA).

    Article Title: New Proof-of-Concept in Viral Inactivation: Virucidal Efficacy of 405 nm Light Against Feline Calicivirus as a Model for Norovirus Decontamination
    Article Snippet: .. Cell and Virus Culture Feline embryonic cells, strain FEA (Jarrett et al. ), were cultured in Dulbecco’s modified eagle’s medium (DMEM) supplemented with 10% foetal bovine serum (FBS), 2 mM l -glutamine, 1 mM sodium pyruvate and 240 U mL−1 penicillin streptomycin (Gibco, Life Technologies, UK), to form 10% FBS-DMEM. ..

    Article Title: The histone deacetylase inhibitor, romidepsin, as a potential treatment for pulmonary fibrosis
    Article Snippet: .. Fibroblasts were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% foetal bovine serum (FBS), 50 units/ml penicillin, 50μg/ml streptomycin, 2mM L-glutamine, 1mM sodium pyruvate and 1x non-essential amino acids (DMEM/FBS) (all from Life Technologies, Paisley, UK). ..

    Article Title: Fibulin1C peptide induces cell attachment and extracellular matrix deposition in lung fibroblasts
    Article Snippet: .. ASM cells and fibroblasts were maintained in Dulbecco's Modified Eagle Medium (DMEM) (Gibco, Grand Island, New York, US) with 5% foetal bovine serum (FBS) and 1% Antibiotic-Antimycotic (Gibco, Grand Island, New York, US). ..

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways
    Article Snippet: .. Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA). .. Calcium chloride dehydrate (CaCl2 ·2H2 O), sodium bicarbonate, hepes, dimethyl sulphoxide (DMSO) and thiazolyl blue tetrazolium bromide (MTT) were from Sigma-Aldrich (St. Louis, MO, USA).

    Article Title: Anti-Oxidative and Anti-Inflammatory Activity of Kenya Grade AA Green Coffee Bean Extracts
    Article Snippet: .. Cell line, Raw 264.7, a macrophage that is used for cell culture and cell toxicity measurement was purchased from Korea cell line bank (Korea Cell Line Bank, Seoul, Korea), and Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), phosphate buffered saline (PBS), penicillin/streptomycin, trypsin for cell culture was purchased from Thermo Scientific HyClone (Logan, UT, USA). .. Haemacytometer (Marienfeld, Germany) and 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), which were used for cell toxicity measurement were purchased from Sigma Chemical Co. (St. Louis, MO, USA), and dimethyl sulfoxide (DMSO) was purchased from Bioshop (Burlington, ON, Canada).

    Western Blot:

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways
    Article Snippet: Reagents Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA). .. Western blots were carried out by the antibodies purchased from Cell Signaling Technology® (Danvers, MA): Estrogen Receptor α (D8H8) Rabbit mAb, Phospho-Estrogen Receptor α (Ser167) (D1A3) Rabbit mAb, p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb, Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb, Akt (pan) (C67E7) Rabbit mAb, Phospho-Akt (Ser473) and GAPDH (14C10) Rabbit mAb.

    Article Title: Four Cysteine Residues Contribute to Homodimerization of Chicken Interleukin-2
    Article Snippet: .. Equal amounts of the chIL-2 dimer were incubated in PBS buffer containing dissociation reagents in a series of five-fold dilutions from the initial concentrations of 4 M KSCN [ ], 10% sodium cholate [ ], 20% Triton X-100 [ ], 8 M urea, 4.8 M guanidine hydrochloride [ ], 215 mM EDTA [ ], 6 M β-ME and 0.8 M DTT, at 37 °C for 1 h. To test the effect of foetal bovine serum (FBS) on dimerization of chIL-2, freshly purified chIL-2 (mostly monomeric) and chIL-2 dimer (post incubation at 37 °C for 24 h) were added to the T cell culture medium (RPMI 1640 medium supplemented with 10% FBS (Gibco, Waltham, MA, USA), 100 U penicillin, and 100 mg streptomycin (Gibco)) and incubated at 37 °C for 24 h. Western blot was employed for the detection of chIL-2. ..

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways
    Article Snippet: Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA). .. Western blots were carried out by the antibodies purchased from Cell Signaling Technology® (Danvers, MA): Estrogen Receptor α (D8H8) Rabbit mAb, Phospho-Estrogen Receptor α (Ser167) (D1A3) Rabbit mAb, p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb, Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb, Akt (pan) (C67E7) Rabbit mAb, Phospho-Akt (Ser473) and GAPDH (14C10) Rabbit mAb.

    Derivative Assay:

    Article Title: Protein kinase D1 regulates ERα-positive breast cancer cell growth response to 17β-estradiol and contributes to poor prognosis in patients
    Article Snippet: Cell culture Breast tissue derived cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) or from the German Resource Centre for Biological Material (DSMZ, Braunschweig, Germany). .. The MCF-7 cell line was cultured in DMEM-Glutamax®, supplemented with 10% foetal bovine serum (FBS) and 100 units/ml penicillin and 100 μg/ml streptomycin (P/S); complete medium (Invitrogen-Life Technologies, Cergy-Pontoise, France).

    Article Title: Fibulin1C peptide induces cell attachment and extracellular matrix deposition in lung fibroblasts
    Article Snippet: Briefly, ASM cells were isolated from medium to large airways and fibroblasts were derived from proximal lung tissues which were obtained from macroscopically normal surgical tailings following resection for thoracic lesions, and from lung transplantation for emphysema or pulmonary fibrosis. .. ASM cells and fibroblasts were maintained in Dulbecco's Modified Eagle Medium (DMEM) (Gibco, Grand Island, New York, US) with 5% foetal bovine serum (FBS) and 1% Antibiotic-Antimycotic (Gibco, Grand Island, New York, US).

    Transfection:

    Article Title: Protein kinase D1 regulates ERα-positive breast cancer cell growth response to 17β-estradiol and contributes to poor prognosis in patients
    Article Snippet: The MCF-7 cell line was cultured in DMEM-Glutamax®, supplemented with 10% foetal bovine serum (FBS) and 100 units/ml penicillin and 100 μg/ml streptomycin (P/S); complete medium (Invitrogen-Life Technologies, Cergy-Pontoise, France). .. 1 mg/ml G418 (Calbiochem, Darmstadt, Germany) was added to this medium for stably transfected MCF-7 cell culture.

    Cell Culture:

    Article Title: Rapid induction of pancreatic cancer cells to cancer stem cells via heterochromatin modulation
    Article Snippet: .. Panc1 and H446 cells were cultured in DMEM (Gibco-life Technologies, USA) containing 10% fetal bovine serum (FBS) (Gibco/Invitrogen, Australia), 1% penicillin/streptomycin (PS) (Invitrogen). .. Bxpc3 and Eca9706 cells were cultured in RPMI-1640 (Gibco-life Technologies, USA) supplemented with 10% FBS and 1% PS.

    Article Title: Rigid Cooperation of Per1 and Per2 proteins
    Article Snippet: .. Cells were cultured in ES medium consisting of Glasgow Minimum Essential Medium (GMEM, Gibco), 10% KnockOut™ Serum Replacement (KSR; Gibco), 1% foetal bovine serum (FBS), 1 mM sodium pyruvate (Gibco), 1 × MEM Non-Essential Amino Acids (Gibco), 100 μM β-mercaptoethanol (Wako, β-ME). .. Then, two inhibitors (2i), 3 μM CHIR99021 (Axon) and 1 μM PD0325901 (Wako) were added.

    Article Title: MicroRNA-138-5p regulates neural stem cell proliferation and differentiation in vitro by targeting TRIP6 expression
    Article Snippet: .. For neural differentiation, NSCs were cultured in an environment with 1 mM retinoic acid (Sigma-Aldrich; Merck KGaA) and 0.5% foetal bovine serum (FBS) for 3 days in 0.5% N2 with Euromed-N medium (Invitrogen; Thermo Fisher Scientific, Inc.) to induce neural differentiation. ..

    Article Title: New Proof-of-Concept in Viral Inactivation: Virucidal Efficacy of 405 nm Light Against Feline Calicivirus as a Model for Norovirus Decontamination
    Article Snippet: .. Cell and Virus Culture Feline embryonic cells, strain FEA (Jarrett et al. ), were cultured in Dulbecco’s modified eagle’s medium (DMEM) supplemented with 10% foetal bovine serum (FBS), 2 mM l -glutamine, 1 mM sodium pyruvate and 240 U mL−1 penicillin streptomycin (Gibco, Life Technologies, UK), to form 10% FBS-DMEM. ..

    Article Title: Protein kinase D1 regulates ERα-positive breast cancer cell growth response to 17β-estradiol and contributes to poor prognosis in patients
    Article Snippet: .. The MCF-7 cell line was cultured in DMEM-Glutamax®, supplemented with 10% foetal bovine serum (FBS) and 100 units/ml penicillin and 100 μg/ml streptomycin (P/S); complete medium (Invitrogen-Life Technologies, Cergy-Pontoise, France). .. 1 mg/ml G418 (Calbiochem, Darmstadt, Germany) was added to this medium for stably transfected MCF-7 cell culture.

    Article Title: The histone deacetylase inhibitor, romidepsin, as a potential treatment for pulmonary fibrosis
    Article Snippet: .. Fibroblasts were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% foetal bovine serum (FBS), 50 units/ml penicillin, 50μg/ml streptomycin, 2mM L-glutamine, 1mM sodium pyruvate and 1x non-essential amino acids (DMEM/FBS) (all from Life Technologies, Paisley, UK). ..

    Article Title: Four Cysteine Residues Contribute to Homodimerization of Chicken Interleukin-2
    Article Snippet: .. Equal amounts of the chIL-2 dimer were incubated in PBS buffer containing dissociation reagents in a series of five-fold dilutions from the initial concentrations of 4 M KSCN [ ], 10% sodium cholate [ ], 20% Triton X-100 [ ], 8 M urea, 4.8 M guanidine hydrochloride [ ], 215 mM EDTA [ ], 6 M β-ME and 0.8 M DTT, at 37 °C for 1 h. To test the effect of foetal bovine serum (FBS) on dimerization of chIL-2, freshly purified chIL-2 (mostly monomeric) and chIL-2 dimer (post incubation at 37 °C for 24 h) were added to the T cell culture medium (RPMI 1640 medium supplemented with 10% FBS (Gibco, Waltham, MA, USA), 100 U penicillin, and 100 mg streptomycin (Gibco)) and incubated at 37 °C for 24 h. Western blot was employed for the detection of chIL-2. ..

    Article Title: Preconditioning of mesenchymal stromal cells with low-intensity ultrasound: influence on chondrogenesis and directed SOX9 signaling pathways
    Article Snippet: .. The inserts were then carefully removed after the solidification of MSC-laden hydrogels which typically ensued within (two constructs per well) were cultured in six-well tissue culture plates (TCP) in DMEM-high glucose medium supplemented with 10% fetal bovine serum (FBS) (Gibco, USA), 100 nM dexamethasone, 50 μg/ml ascorbic acid, and 1× antibiotic-antimycotic solution (Gibco, USA) at 37 °C and 5% CO2 with or without cLIUS for 6 weeks. .. cLIUS treatment regimen of 3D MSC constructs Six-well TCPs containing MSC-hydrogel constructs were placed in plate holders of a cLIUS-assisted incubator developed at the Department of Chemical Engineering, University of Nebraska-Lincoln (UNL), USA, with operating procedures detailed elsewhere [ ]. cLIUS was applied to the plates at 5 MHz frequency and 2.5 Vpp (14 kPa) for 20 min per application at four applications/day for a period of 6 weeks.

    Article Title: Characterization and Expression of Senescence Marker in Prolonged Passages of Rat Bone Marrow-Derived Mesenchymal Stem Cells
    Article Snippet: Harvested cells were cultured in 25 cm2 flasks for further expansion. .. Media used in rBM-MSC expansion were LDMEM with GLUTAMAX-I (Gibco, United Kingdom), supplemented with 20% foetal bovine serum (FBS), 1% penicillin/streptomycin, 0.5% Fungizone, and 0.1% gentamycin (Gibco, United Kingdom).

    Article Title: Fibulin1C peptide induces cell attachment and extracellular matrix deposition in lung fibroblasts
    Article Snippet: Paragraph title: Study subjects and primary lung cell culture ... ASM cells and fibroblasts were maintained in Dulbecco's Modified Eagle Medium (DMEM) (Gibco, Grand Island, New York, US) with 5% foetal bovine serum (FBS) and 1% Antibiotic-Antimycotic (Gibco, Grand Island, New York, US).

    Article Title: Anti-Oxidative and Anti-Inflammatory Activity of Kenya Grade AA Green Coffee Bean Extracts
    Article Snippet: .. Cell line, Raw 264.7, a macrophage that is used for cell culture and cell toxicity measurement was purchased from Korea cell line bank (Korea Cell Line Bank, Seoul, Korea), and Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), phosphate buffered saline (PBS), penicillin/streptomycin, trypsin for cell culture was purchased from Thermo Scientific HyClone (Logan, UT, USA). .. Haemacytometer (Marienfeld, Germany) and 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), which were used for cell toxicity measurement were purchased from Sigma Chemical Co. (St. Louis, MO, USA), and dimethyl sulfoxide (DMSO) was purchased from Bioshop (Burlington, ON, Canada).

    Generated:

    Article Title: Rigid Cooperation of Per1 and Per2 proteins
    Article Snippet: Per2 (−/−) ES cells were established by Naoshi Koide (RIKEN) from mice generated in this study. .. Cells were cultured in ES medium consisting of Glasgow Minimum Essential Medium (GMEM, Gibco), 10% KnockOut™ Serum Replacement (KSR; Gibco), 1% foetal bovine serum (FBS), 1 mM sodium pyruvate (Gibco), 1 × MEM Non-Essential Amino Acids (Gibco), 100 μM β-mercaptoethanol (Wako, β-ME).

    MTT Assay:

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways
    Article Snippet: Reagents Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA). .. Calcium chloride dehydrate (CaCl2 ·2H2 O), sodium bicarbonate, hepes, dimethyl sulphoxide (DMSO) and thiazolyl blue tetrazolium bromide (MTT) were from Sigma-Aldrich (St. Louis, MO, USA).

    Article Title: siRNAs Targeting Growth Factor Receptor and Anti-Apoptotic Genes Synergistically Kill Breast Cancer Cells through Inhibition of MAPK and PI-3 Kinase Pathways
    Article Snippet: Dulbecco’s modified Eagle medium (DMEM), DMEM powder, foetal bovine serum (FBS), TrypLE Express enzyme (1×) (trypsin-EDTA) and penicillin/streptomycin were obtained from Gibco BRL (Carlsbad, CA, USA). .. Calcium chloride dehydrate (CaCl2 ·2H2 O), sodium bicarbonate, hepes, dimethyl sulphoxide (DMSO) and thiazolyl blue tetrazolium bromide (MTT) were from Sigma-Aldrich (St. Louis, MO, USA).

    Article Title: Anti-Oxidative and Anti-Inflammatory Activity of Kenya Grade AA Green Coffee Bean Extracts
    Article Snippet: Cell line, Raw 264.7, a macrophage that is used for cell culture and cell toxicity measurement was purchased from Korea cell line bank (Korea Cell Line Bank, Seoul, Korea), and Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), phosphate buffered saline (PBS), penicillin/streptomycin, trypsin for cell culture was purchased from Thermo Scientific HyClone (Logan, UT, USA). .. Haemacytometer (Marienfeld, Germany) and 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), which were used for cell toxicity measurement were purchased from Sigma Chemical Co. (St. Louis, MO, USA), and dimethyl sulfoxide (DMSO) was purchased from Bioshop (Burlington, ON, Canada).

    Multiple Displacement Amplification:

    Article Title: Protein kinase D1 regulates ERα-positive breast cancer cell growth response to 17β-estradiol and contributes to poor prognosis in patients
    Article Snippet: For the other in vitro experiments, MCF-7 and MDA-MB-415 cell lines were cultured as follows. .. The MCF-7 cell line was cultured in DMEM-Glutamax®, supplemented with 10% foetal bovine serum (FBS) and 100 units/ml penicillin and 100 μg/ml streptomycin (P/S); complete medium (Invitrogen-Life Technologies, Cergy-Pontoise, France).

    Isolation:

    Article Title: Fibulin1C peptide induces cell attachment and extracellular matrix deposition in lung fibroblasts
    Article Snippet: Briefly, ASM cells were isolated from medium to large airways and fibroblasts were derived from proximal lung tissues which were obtained from macroscopically normal surgical tailings following resection for thoracic lesions, and from lung transplantation for emphysema or pulmonary fibrosis. .. ASM cells and fibroblasts were maintained in Dulbecco's Modified Eagle Medium (DMEM) (Gibco, Grand Island, New York, US) with 5% foetal bovine serum (FBS) and 1% Antibiotic-Antimycotic (Gibco, Grand Island, New York, US).

    Mouse Assay:

    Article Title: Rigid Cooperation of Per1 and Per2 proteins
    Article Snippet: Per2 (−/−) ES cells were established by Naoshi Koide (RIKEN) from mice generated in this study. .. Cells were cultured in ES medium consisting of Glasgow Minimum Essential Medium (GMEM, Gibco), 10% KnockOut™ Serum Replacement (KSR; Gibco), 1% foetal bovine serum (FBS), 1 mM sodium pyruvate (Gibco), 1 × MEM Non-Essential Amino Acids (Gibco), 100 μM β-mercaptoethanol (Wako, β-ME).

    Purification:

    Article Title: Four Cysteine Residues Contribute to Homodimerization of Chicken Interleukin-2
    Article Snippet: .. Equal amounts of the chIL-2 dimer were incubated in PBS buffer containing dissociation reagents in a series of five-fold dilutions from the initial concentrations of 4 M KSCN [ ], 10% sodium cholate [ ], 20% Triton X-100 [ ], 8 M urea, 4.8 M guanidine hydrochloride [ ], 215 mM EDTA [ ], 6 M β-ME and 0.8 M DTT, at 37 °C for 1 h. To test the effect of foetal bovine serum (FBS) on dimerization of chIL-2, freshly purified chIL-2 (mostly monomeric) and chIL-2 dimer (post incubation at 37 °C for 24 h) were added to the T cell culture medium (RPMI 1640 medium supplemented with 10% FBS (Gibco, Waltham, MA, USA), 100 U penicillin, and 100 mg streptomycin (Gibco)) and incubated at 37 °C for 24 h. Western blot was employed for the detection of chIL-2. ..

    Plasmid Preparation:

    Article Title: De novo lipogenesis represents a therapeutic target in mutant Kras non-small cell lung cancer
    Article Snippet: H858 and H1299 cells were transduced with green fluorescent protein (GFP) vector control or retrovirus expressing KrasG12V . .. For cell proliferation, cells were seeded into 6- or 24-well plates overnight and then treated with the indicated doses of inhibitors using Roswell Park Memorial Institute (RPMI)-1640 (Buffalo, NY, USA) medium supplemented with either 1% fetal bovine serum (FBS), and the number of cells was determined after 3–5 d by hemocytometer or Countess Automated Cell Counter (Thermo Fisher Scientific, Waltham, MA, USA).

    shRNA:

    Article Title: De novo lipogenesis represents a therapeutic target in mutant Kras non-small cell lung cancer
    Article Snippet: A549 and H23 NSCLC cells were transduced with lentivirus expressing nontarget short hairpin RNA (shRNA) pLKO (A549NTshRNA and H23NTshRNA ) or shRNA against Kras (A549sh Kras and H23sh Kras ; TRCN0000033260,TRCN0000033262, TRCN0000033261, and TRCN0000033259; MilliporeSigma, Burlington, MA, USA). .. For cell proliferation, cells were seeded into 6- or 24-well plates overnight and then treated with the indicated doses of inhibitors using Roswell Park Memorial Institute (RPMI)-1640 (Buffalo, NY, USA) medium supplemented with either 1% fetal bovine serum (FBS), and the number of cells was determined after 3–5 d by hemocytometer or Countess Automated Cell Counter (Thermo Fisher Scientific, Waltham, MA, USA).

    In Vitro:

    Article Title: Protein kinase D1 regulates ERα-positive breast cancer cell growth response to 17β-estradiol and contributes to poor prognosis in patients
    Article Snippet: For the other in vitro experiments, MCF-7 and MDA-MB-415 cell lines were cultured as follows. .. The MCF-7 cell line was cultured in DMEM-Glutamax®, supplemented with 10% foetal bovine serum (FBS) and 100 units/ml penicillin and 100 μg/ml streptomycin (P/S); complete medium (Invitrogen-Life Technologies, Cergy-Pontoise, France).

    Article Title: De novo lipogenesis represents a therapeutic target in mutant Kras non-small cell lung cancer
    Article Snippet: Paragraph title: Genetic manipulation of Kras in vitro ... For cell proliferation, cells were seeded into 6- or 24-well plates overnight and then treated with the indicated doses of inhibitors using Roswell Park Memorial Institute (RPMI)-1640 (Buffalo, NY, USA) medium supplemented with either 1% fetal bovine serum (FBS), and the number of cells was determined after 3–5 d by hemocytometer or Countess Automated Cell Counter (Thermo Fisher Scientific, Waltham, MA, USA).

    Knock-Out:

    Article Title: Rigid Cooperation of Per1 and Per2 proteins
    Article Snippet: .. Cells were cultured in ES medium consisting of Glasgow Minimum Essential Medium (GMEM, Gibco), 10% KnockOut™ Serum Replacement (KSR; Gibco), 1% foetal bovine serum (FBS), 1 mM sodium pyruvate (Gibco), 1 × MEM Non-Essential Amino Acids (Gibco), 100 μM β-mercaptoethanol (Wako, β-ME). .. Then, two inhibitors (2i), 3 μM CHIR99021 (Axon) and 1 μM PD0325901 (Wako) were added.

    Concentration Assay:

    Article Title: The interaction of fluorescent nanodiamond probes with cellular media
    Article Snippet: Aggregates were created by dispersing the FNDs25 to an end concentration of 200 μg mL−1 in different media. .. The different media were (1) DMEM Complete (consisting of DMEM + complements: Glutamax (1%), Pen/Strep (1%), Foetal bovine serum (FBS) (10%), Gibco Life Technologies, Bleiswijk, the Netherlands, www.thermofisher.com/ch/en/home/brands/gibco ). (2) DMEM without complements and (3) pure or diluted (10%) FBS.

    Article Title: Rigid Cooperation of Per1 and Per2 proteins
    Article Snippet: Cells were cultured in ES medium consisting of Glasgow Minimum Essential Medium (GMEM, Gibco), 10% KnockOut™ Serum Replacement (KSR; Gibco), 1% foetal bovine serum (FBS), 1 mM sodium pyruvate (Gibco), 1 × MEM Non-Essential Amino Acids (Gibco), 100 μM β-mercaptoethanol (Wako, β-ME). .. ESGRO mouse leukaemia inhibitory factor (mLIF, Millipore) was added to a final concentration of 2 × 103 U/mL.

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  • 99
    Thermo Fisher mcp 1
    Enhanced chemoattractant activity in CAV-1-silenced monocytes. The chemoattractant activity of monocytes toward <t>MCP-1</t> was assessed by chemotaxis assay. Cav-1-silenced (siCAV-1) or control monocytes (siNeg) were seeded in the upper compartment, and the medium containing 100 ng/ml MCP-1 was then added to the lower compartment. After a 90-minute incubation, cells were collected from both compartments and manually counted. Graph displays the ratio of migrating monocytes. Monocytes were obtained from three donors, and results represented three independent experiments. Data are shown as a mean ± SD. One-way analysis of variance (post hoc Tukey), ***P
    Mcp 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 283 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mcp 1/product/Thermo Fisher
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    85
    Thermo Fisher hap1 uptake medium
    TSSC1 KO impairs STxB retrograde transport. (A) Live WT and TSSC1-KO <t>HAP1</t> cells, and TSSC1-KO HAP1 cells rescued by stable expression of TSSC1-FOS, were incubated with Cy3-STxB for 1 h, fixed, stained for the Golgi marker giantin, and examined by confocal fluorescence microscopy. Scale bar, 10 μm. (B) Spearman’s rank coefficient quantification of data set in A, expressed as mean; error bars, SEM. Minimum of nine cells. *** p ≤ 0.001.
    Hap1 Uptake Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher ocum 2md3 cells
    Antiproliferative effects of eribulin and 5-FU on gastric cancer cell lines and HPMCs. Notes: Proliferation in the presence of eribulin or 5-FU was measured by MTT assay. Values shown are means ± SD of three experiments. Respective IC 50 values of eribulin and 5-FU were 2.2 ± 0.6 and 9.7 ± 3.7 μM for MKN-45 cells ( A ), 0.7 ± 0.2 and 9.2 ± 0.9 μM for MKN-74 cells ( B ), 0.5 ± 0.1 and 4.9 ± 1.8 μM for MKN-7 cells ( C ), 1.7 ± 0.4 and 9.1 ± 0.2 μM for <t>OCUM-2MD3</t> cells ( D ), and 8.1 ± 0.6 and 17.9 ± 6.4 μM for HPMCs ( E ). Abbreviations: 5-FU, 5-fluorouracil; HPMCs, human peritoneal mesothelial cells; IC 50 , half maximal inhibitory concentration; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.
    Ocum 2md3 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Enhanced chemoattractant activity in CAV-1-silenced monocytes. The chemoattractant activity of monocytes toward MCP-1 was assessed by chemotaxis assay. Cav-1-silenced (siCAV-1) or control monocytes (siNeg) were seeded in the upper compartment, and the medium containing 100 ng/ml MCP-1 was then added to the lower compartment. After a 90-minute incubation, cells were collected from both compartments and manually counted. Graph displays the ratio of migrating monocytes. Monocytes were obtained from three donors, and results represented three independent experiments. Data are shown as a mean ± SD. One-way analysis of variance (post hoc Tukey), ***P

    Journal: Scientific Reports

    Article Title: Downregulated Caveolin-1 expression in circulating monocytes may contribute to the pathogenesis of psoriasis

    doi: 10.1038/s41598-018-36767-5

    Figure Lengend Snippet: Enhanced chemoattractant activity in CAV-1-silenced monocytes. The chemoattractant activity of monocytes toward MCP-1 was assessed by chemotaxis assay. Cav-1-silenced (siCAV-1) or control monocytes (siNeg) were seeded in the upper compartment, and the medium containing 100 ng/ml MCP-1 was then added to the lower compartment. After a 90-minute incubation, cells were collected from both compartments and manually counted. Graph displays the ratio of migrating monocytes. Monocytes were obtained from three donors, and results represented three independent experiments. Data are shown as a mean ± SD. One-way analysis of variance (post hoc Tukey), ***P

    Article Snippet: RPMI-1640 supplemented with 1% FBS in either the presence or absence of 100 ng/ml MCP-1 (Thermo Fisher Scientific) was added into the lower compartment.

    Techniques: Activity Assay, Chemotaxis Assay, Incubation

    TSSC1 KO impairs STxB retrograde transport. (A) Live WT and TSSC1-KO HAP1 cells, and TSSC1-KO HAP1 cells rescued by stable expression of TSSC1-FOS, were incubated with Cy3-STxB for 1 h, fixed, stained for the Golgi marker giantin, and examined by confocal fluorescence microscopy. Scale bar, 10 μm. (B) Spearman’s rank coefficient quantification of data set in A, expressed as mean; error bars, SEM. Minimum of nine cells. *** p ≤ 0.001.

    Journal: Molecular Biology of the Cell

    Article Title: TSSC1 is novel component of the endosomal retrieval machinery

    doi: 10.1091/mbc.E16-04-0209

    Figure Lengend Snippet: TSSC1 KO impairs STxB retrograde transport. (A) Live WT and TSSC1-KO HAP1 cells, and TSSC1-KO HAP1 cells rescued by stable expression of TSSC1-FOS, were incubated with Cy3-STxB for 1 h, fixed, stained for the Golgi marker giantin, and examined by confocal fluorescence microscopy. Scale bar, 10 μm. (B) Spearman’s rank coefficient quantification of data set in A, expressed as mean; error bars, SEM. Minimum of nine cells. *** p ≤ 0.001.

    Article Snippet: In brief, cells were incubated for 30 min in HAP1 uptake medium (Iscove’s modified Dulbecco’s medium [IMDM; ThermoFisher Scientific] –FBS + 1% BSA).

    Techniques: Expressing, Incubation, Staining, Marker, Fluorescence, Microscopy

    Biochemical analysis of TSSC1. (A–C) Extracts from H4 cells stably expressing BLOS2-OSF (negative control), Syndetin-OSF (positive control), or TSSC1-FOS were analyzed by pull down with StrepTactin beads, followed by SDS–PAGE and immunoblotting for coexpressed ANG2-13myc and endogenous VPS52, VPS53, and actin (negative control). (D) Extracts from untransfected and stably expressing GFP or VPS54-GFP cells were analyzed by pull down with a nanobody to GFP conjugated to magnetic beads, followed by SDS–PAGE and immunoblotting for endogenous TSSC1 and actin (negative control). *Nonspecific band. (E) WT and TSSC1-KO HAP1 cells, and TSSC1-KO HAP1 cells rescued by stable expression of TSSC1-FOS, were analyzed by SDS–PAGE and immunoblotting with antibodies to TSSC1, ANG2, VPS52, VPS53, Syndetin, or actin (negative control). (F) SDS–PAGE and immunoblot analysis of the expression of Syndetin, ANG2, TSSC1, and actin in multiple rat tissues. (G) A rat brain extract was subjected to immunoprecipitation with antibodies to immunoglobulin G (negative control) or TSSC1 followed by SDS–PAGE and immunoblot analysis of the precipitates with antibodies to ANG2, VPS52, VPS53, Syndetin, and actin. A 1% fraction of the input extract was run alongside for comparison. (H) Gel filtration of rat brain extract on a Superose 6 column and analysis of the fractions indicated on top by SDS–PAGE and immunoblotting with antibodies to ANG2 and TSSC1. In all the immunoblots, the positions of molecular mass markers (in kilodaltons) are indicated on the left.

    Journal: Molecular Biology of the Cell

    Article Title: TSSC1 is novel component of the endosomal retrieval machinery

    doi: 10.1091/mbc.E16-04-0209

    Figure Lengend Snippet: Biochemical analysis of TSSC1. (A–C) Extracts from H4 cells stably expressing BLOS2-OSF (negative control), Syndetin-OSF (positive control), or TSSC1-FOS were analyzed by pull down with StrepTactin beads, followed by SDS–PAGE and immunoblotting for coexpressed ANG2-13myc and endogenous VPS52, VPS53, and actin (negative control). (D) Extracts from untransfected and stably expressing GFP or VPS54-GFP cells were analyzed by pull down with a nanobody to GFP conjugated to magnetic beads, followed by SDS–PAGE and immunoblotting for endogenous TSSC1 and actin (negative control). *Nonspecific band. (E) WT and TSSC1-KO HAP1 cells, and TSSC1-KO HAP1 cells rescued by stable expression of TSSC1-FOS, were analyzed by SDS–PAGE and immunoblotting with antibodies to TSSC1, ANG2, VPS52, VPS53, Syndetin, or actin (negative control). (F) SDS–PAGE and immunoblot analysis of the expression of Syndetin, ANG2, TSSC1, and actin in multiple rat tissues. (G) A rat brain extract was subjected to immunoprecipitation with antibodies to immunoglobulin G (negative control) or TSSC1 followed by SDS–PAGE and immunoblot analysis of the precipitates with antibodies to ANG2, VPS52, VPS53, Syndetin, and actin. A 1% fraction of the input extract was run alongside for comparison. (H) Gel filtration of rat brain extract on a Superose 6 column and analysis of the fractions indicated on top by SDS–PAGE and immunoblotting with antibodies to ANG2 and TSSC1. In all the immunoblots, the positions of molecular mass markers (in kilodaltons) are indicated on the left.

    Article Snippet: In brief, cells were incubated for 30 min in HAP1 uptake medium (Iscove’s modified Dulbecco’s medium [IMDM; ThermoFisher Scientific] –FBS + 1% BSA).

    Techniques: Stable Transfection, Expressing, Negative Control, Positive Control, SDS Page, Magnetic Beads, Immunoprecipitation, Filtration, Western Blot

    (A) Live-cell imaging of Cy3-STxB uptake in WT, VPS54-KO, and TSSC1-KO HAP1 cells. Cells were incubated for 1 h in the continuous presence of Cy3-STxB and imaged over the course of 1½ h by spinning-disk confocal microscopy. At each time point, a z -stack of the whole cell was taken and averaged into a maximum intensity projection. Scale bar, 10 μm. See Supplemental Videos S1–S3 for complete time course. (B) Distance quantification of data set from A. Each point plotted represents an individual detectable punctate structure, plotted showing its distance from the center of the nucleus. Left, combination of at least 70 data points from multiple cells over time; far right, combination of all data, demonstrating the relative proximity of the STxB punctae to the nucleus in the WT cells. Significance was calculated using Student’s t test. (C) FACS analysis of Alexa 488–Tf uptake into control HeLa cells or HeLa cells treated with siRNAs to TSSC1 or Syndetin. The plot represents the geometric mean of Alexa 488–Tf intensity of the population (∼30,000 cells) as a function of time. Error is the SEM from multiple independent experiments. Left, continuous exposure to Alexa 488–Tf (significance calculated as two-way analysis of variance); right, chase experiment after 30-min uptake (significance calculated using Student’s t test at selected time points). ns: p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

    Journal: Molecular Biology of the Cell

    Article Title: TSSC1 is novel component of the endosomal retrieval machinery

    doi: 10.1091/mbc.E16-04-0209

    Figure Lengend Snippet: (A) Live-cell imaging of Cy3-STxB uptake in WT, VPS54-KO, and TSSC1-KO HAP1 cells. Cells were incubated for 1 h in the continuous presence of Cy3-STxB and imaged over the course of 1½ h by spinning-disk confocal microscopy. At each time point, a z -stack of the whole cell was taken and averaged into a maximum intensity projection. Scale bar, 10 μm. See Supplemental Videos S1–S3 for complete time course. (B) Distance quantification of data set from A. Each point plotted represents an individual detectable punctate structure, plotted showing its distance from the center of the nucleus. Left, combination of at least 70 data points from multiple cells over time; far right, combination of all data, demonstrating the relative proximity of the STxB punctae to the nucleus in the WT cells. Significance was calculated using Student’s t test. (C) FACS analysis of Alexa 488–Tf uptake into control HeLa cells or HeLa cells treated with siRNAs to TSSC1 or Syndetin. The plot represents the geometric mean of Alexa 488–Tf intensity of the population (∼30,000 cells) as a function of time. Error is the SEM from multiple independent experiments. Left, continuous exposure to Alexa 488–Tf (significance calculated as two-way analysis of variance); right, chase experiment after 30-min uptake (significance calculated using Student’s t test at selected time points). ns: p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

    Article Snippet: In brief, cells were incubated for 30 min in HAP1 uptake medium (Iscove’s modified Dulbecco’s medium [IMDM; ThermoFisher Scientific] –FBS + 1% BSA).

    Techniques: Live Cell Imaging, Incubation, Confocal Microscopy, FACS

    Antiproliferative effects of eribulin and 5-FU on gastric cancer cell lines and HPMCs. Notes: Proliferation in the presence of eribulin or 5-FU was measured by MTT assay. Values shown are means ± SD of three experiments. Respective IC 50 values of eribulin and 5-FU were 2.2 ± 0.6 and 9.7 ± 3.7 μM for MKN-45 cells ( A ), 0.7 ± 0.2 and 9.2 ± 0.9 μM for MKN-74 cells ( B ), 0.5 ± 0.1 and 4.9 ± 1.8 μM for MKN-7 cells ( C ), 1.7 ± 0.4 and 9.1 ± 0.2 μM for OCUM-2MD3 cells ( D ), and 8.1 ± 0.6 and 17.9 ± 6.4 μM for HPMCs ( E ). Abbreviations: 5-FU, 5-fluorouracil; HPMCs, human peritoneal mesothelial cells; IC 50 , half maximal inhibitory concentration; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.

    Journal: Cancer Management and Research

    Article Title: Low-dose eribulin mesylate exerts antitumor effects in gastric cancer by inhibiting fibrosis via the suppression of epithelial–mesenchymal transition and acts synergistically with 5-fluorouracil

    doi: 10.2147/CMAR.S167846

    Figure Lengend Snippet: Antiproliferative effects of eribulin and 5-FU on gastric cancer cell lines and HPMCs. Notes: Proliferation in the presence of eribulin or 5-FU was measured by MTT assay. Values shown are means ± SD of three experiments. Respective IC 50 values of eribulin and 5-FU were 2.2 ± 0.6 and 9.7 ± 3.7 μM for MKN-45 cells ( A ), 0.7 ± 0.2 and 9.2 ± 0.9 μM for MKN-74 cells ( B ), 0.5 ± 0.1 and 4.9 ± 1.8 μM for MKN-7 cells ( C ), 1.7 ± 0.4 and 9.1 ± 0.2 μM for OCUM-2MD3 cells ( D ), and 8.1 ± 0.6 and 17.9 ± 6.4 μM for HPMCs ( E ). Abbreviations: 5-FU, 5-fluorouracil; HPMCs, human peritoneal mesothelial cells; IC 50 , half maximal inhibitory concentration; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.

    Article Snippet: MKN-45, MKN-74, and MKN-7 cells were maintained in RPMI-1640 medium supplemented with 10% FBS, and OCUM-2MD3 cells were cultured in Dulbecco’s modified Eagle’s medium (Thermo Fisher Scientific) containing 10% FBS.

    Techniques: MTT Assay, Concentration Assay