dulbecco modified minimum essential medium  (Thermo Fisher)


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    Name:
    Dulbecco s Modified Eagle s Limiting Medium DMEM LM
    Description:
    Formulation Sterile filtered DMEM L leucine L methionine with 4 5g L glucose 4 0mM L glutamine sodium pyruvate and phenol red Related Products L Photo Leucine L Photo Methionine
    Catalog Number:
    30030
    Price:
    None
    Category:
    Labeling Detection Products
    Applications:
    Protein Biology|Protein Crosslinking|Protein Labeling & Crosslinking
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    Structured Review

    Thermo Fisher dulbecco modified minimum essential medium
    Studies of West Nile virus (WNV) properties in cell cultures and mice. A) Plaque morphology of WNV NY99 , prototype WNV KUN, and WNV NSW2011 in Vero cells. Cells in 6-well plates were infected with specified virus and overlaid with 0.75% low melting point agarose in <t>Dulbecco</t> modified minimum essential medium (Life Technologies, Carlsbad, CA, USA) containing 2% fetal bovine serum. Four days after infection, the cells were fixed with 4% formaldehyde and stained with 0.2% crystal violet. B) Assessment of envelope (E) protein glycosylation of WNV NSW2011 , WNV KUN and WNV NY99 by endoglycosidase digestion (PNGase F; Roche Diagnostics, Basel, Switzerland). Viral proteins in culture supernatant were digested by PNGase F (+) or undigested (−) and then resolved on sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The migration rate of the E protein in each sample was determined by Western blot with E glycoprotein–specific monoclonal antibodies. C) Young adult (4 weeks old) or D) weanling (18–19 days old) Swiss outbred mice survival after intraperitoneal injection with 1,000 PFU (adult) or 10 PFU (weanling) of WNV NY99 , WNV KUN , or WNV NSW2011 . The mice were monitored for 21 days after injection for signs of encephalitis and then euthanized. The differences in virulence in weanling and adult mice between different pairs of viruses were all highly significant, as calculated by log rank Mantel-Cox algorithm with exact p values: for adult mice, WNV NY99 vs. WNV KUN p
    Formulation Sterile filtered DMEM L leucine L methionine with 4 5g L glucose 4 0mM L glutamine sodium pyruvate and phenol red Related Products L Photo Leucine L Photo Methionine
    https://www.bioz.com/result/dulbecco modified minimum essential medium/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
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    dulbecco modified minimum essential medium - by Bioz Stars, 2021-06
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    Images

    1) Product Images from "Characterization of Virulent West Nile Virus Kunjin Strain, Australia, 2011"

    Article Title: Characterization of Virulent West Nile Virus Kunjin Strain, Australia, 2011

    Journal: Emerging Infectious Diseases

    doi: 10.3201/eid1805.111720

    Studies of West Nile virus (WNV) properties in cell cultures and mice. A) Plaque morphology of WNV NY99 , prototype WNV KUN, and WNV NSW2011 in Vero cells. Cells in 6-well plates were infected with specified virus and overlaid with 0.75% low melting point agarose in Dulbecco modified minimum essential medium (Life Technologies, Carlsbad, CA, USA) containing 2% fetal bovine serum. Four days after infection, the cells were fixed with 4% formaldehyde and stained with 0.2% crystal violet. B) Assessment of envelope (E) protein glycosylation of WNV NSW2011 , WNV KUN and WNV NY99 by endoglycosidase digestion (PNGase F; Roche Diagnostics, Basel, Switzerland). Viral proteins in culture supernatant were digested by PNGase F (+) or undigested (−) and then resolved on sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The migration rate of the E protein in each sample was determined by Western blot with E glycoprotein–specific monoclonal antibodies. C) Young adult (4 weeks old) or D) weanling (18–19 days old) Swiss outbred mice survival after intraperitoneal injection with 1,000 PFU (adult) or 10 PFU (weanling) of WNV NY99 , WNV KUN , or WNV NSW2011 . The mice were monitored for 21 days after injection for signs of encephalitis and then euthanized. The differences in virulence in weanling and adult mice between different pairs of viruses were all highly significant, as calculated by log rank Mantel-Cox algorithm with exact p values: for adult mice, WNV NY99 vs. WNV KUN p
    Figure Legend Snippet: Studies of West Nile virus (WNV) properties in cell cultures and mice. A) Plaque morphology of WNV NY99 , prototype WNV KUN, and WNV NSW2011 in Vero cells. Cells in 6-well plates were infected with specified virus and overlaid with 0.75% low melting point agarose in Dulbecco modified minimum essential medium (Life Technologies, Carlsbad, CA, USA) containing 2% fetal bovine serum. Four days after infection, the cells were fixed with 4% formaldehyde and stained with 0.2% crystal violet. B) Assessment of envelope (E) protein glycosylation of WNV NSW2011 , WNV KUN and WNV NY99 by endoglycosidase digestion (PNGase F; Roche Diagnostics, Basel, Switzerland). Viral proteins in culture supernatant were digested by PNGase F (+) or undigested (−) and then resolved on sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The migration rate of the E protein in each sample was determined by Western blot with E glycoprotein–specific monoclonal antibodies. C) Young adult (4 weeks old) or D) weanling (18–19 days old) Swiss outbred mice survival after intraperitoneal injection with 1,000 PFU (adult) or 10 PFU (weanling) of WNV NY99 , WNV KUN , or WNV NSW2011 . The mice were monitored for 21 days after injection for signs of encephalitis and then euthanized. The differences in virulence in weanling and adult mice between different pairs of viruses were all highly significant, as calculated by log rank Mantel-Cox algorithm with exact p values: for adult mice, WNV NY99 vs. WNV KUN p

    Techniques Used: Mouse Assay, Infection, Modification, Staining, Polyacrylamide Gel Electrophoresis, Migration, Western Blot, Injection

    2) Product Images from "Neuropeptide Y, substance P, and human bone morphogenetic protein 2 stimulate human osteoblast osteogenic activity by enhancing gap junction intercellular communication"

    Article Title: Neuropeptide Y, substance P, and human bone morphogenetic protein 2 stimulate human osteoblast osteogenic activity by enhancing gap junction intercellular communication

    Journal: Brazilian Journal of Medical and Biological Research

    doi: 10.1590/1414-431X20144226

    Cellular morphology of human osteoblasts. The osteoblast was thawed by directly putting the freezing tube into water at 40°C. Then, the frozen solution was removed by centrifugation (200 g for 1 min). The cells were cultured in Dulbecco's Modified Eagle Medium supplemented with 10% fetal bovine serum. The phase-contrast microscopic images of the cells showed that osteoblasts were fusiform or polygonal, with plenty of cytoplasm and large oval nuclei ( A ). SEM showed that the cells were connected by ecphyma ( B ).
    Figure Legend Snippet: Cellular morphology of human osteoblasts. The osteoblast was thawed by directly putting the freezing tube into water at 40°C. Then, the frozen solution was removed by centrifugation (200 g for 1 min). The cells were cultured in Dulbecco's Modified Eagle Medium supplemented with 10% fetal bovine serum. The phase-contrast microscopic images of the cells showed that osteoblasts were fusiform or polygonal, with plenty of cytoplasm and large oval nuclei ( A ). SEM showed that the cells were connected by ecphyma ( B ).

    Techniques Used: Centrifugation, Cell Culture, Modification

    3) Product Images from "Inhibitory Effects of Enalaprilat on Rat Cardiac Fibroblast Proliferation via ROS/P38MAPK/TGF-β1 Signaling Pathway"

    Article Title: Inhibitory Effects of Enalaprilat on Rat Cardiac Fibroblast Proliferation via ROS/P38MAPK/TGF-β1 Signaling Pathway

    Journal: Molecules

    doi: 10.3390/molecules17032738

    Effects of Ena. and antioxidant NAC on ROS intensity of cardiac fibroblasts in fluorescence microscope (200×). CFb were cultured in Dulbecco’s Modified Eagle Medium containing Ang II (10 −7 M) for 10 minutes, 20 minutes and 30 minutes, and treated with enalaprilat (10 −7 M, 10 −6 M, and 10 −5 M) for 30 minutes. CFb were labeled with 2',7'-dichlorofluorescein diacetate for 30 minutes and ROS generation was analyzed by fluorescence detection with fluorescent microscopy at 200×. Upper part: A :CFb were cultured in Dulbecco’s Modified Eagle Medium without Ang II; B : CFb were stimulated with Ang II for 10 min; C : CFb were stimulated with Ang II for 20 min; D : CFb were stimulated with Ang II for30 minutes; E – G : CFb were stimulated with Ang II and treated with Ena.(10 –7 M, 10 –6 M, and 10 –5 M respectively) for 30 minutes; H : CFb were stimulated with Ang II and treated with NAC (10 –2 M, 30 minutes prior to Ang IIadministration) for 30 minutes. Lower part: a and b : Statistic representations are indicated as bars in involved group. Data are presented as mean ± S.E.M. # P
    Figure Legend Snippet: Effects of Ena. and antioxidant NAC on ROS intensity of cardiac fibroblasts in fluorescence microscope (200×). CFb were cultured in Dulbecco’s Modified Eagle Medium containing Ang II (10 −7 M) for 10 minutes, 20 minutes and 30 minutes, and treated with enalaprilat (10 −7 M, 10 −6 M, and 10 −5 M) for 30 minutes. CFb were labeled with 2',7'-dichlorofluorescein diacetate for 30 minutes and ROS generation was analyzed by fluorescence detection with fluorescent microscopy at 200×. Upper part: A :CFb were cultured in Dulbecco’s Modified Eagle Medium without Ang II; B : CFb were stimulated with Ang II for 10 min; C : CFb were stimulated with Ang II for 20 min; D : CFb were stimulated with Ang II for30 minutes; E – G : CFb were stimulated with Ang II and treated with Ena.(10 –7 M, 10 –6 M, and 10 –5 M respectively) for 30 minutes; H : CFb were stimulated with Ang II and treated with NAC (10 –2 M, 30 minutes prior to Ang IIadministration) for 30 minutes. Lower part: a and b : Statistic representations are indicated as bars in involved group. Data are presented as mean ± S.E.M. # P

    Techniques Used: Fluorescence, Microscopy, Cell Culture, Modification, Labeling

    Effects of Ang II, Ena., NAC on p-p38MAPK nuclear translocation. A : p-p38MAPK nuclear translocation was observed under inverted microscope (400×) with immunocytochemistry staining; B : Statistic representation of p-p38MAPK nuclear translocation is indicated as bars in the involved group. CFb were cultured in Dulbecco’s Modified Eagle Medium containing Ang II (10 –7 M). Brown staining in cytoplasm and nuclei represented protein expression of p-p38MAPK. Figure 4 A, A : Control group; B – F : Ang II induced for 2 minutes–30 minutes; G – H : Ena. (10 –6 M) treated for 5 and 15 minutes; I : NAC (10 –2 M, 30 minutes prior to AngII administration) for 15 min. # P
    Figure Legend Snippet: Effects of Ang II, Ena., NAC on p-p38MAPK nuclear translocation. A : p-p38MAPK nuclear translocation was observed under inverted microscope (400×) with immunocytochemistry staining; B : Statistic representation of p-p38MAPK nuclear translocation is indicated as bars in the involved group. CFb were cultured in Dulbecco’s Modified Eagle Medium containing Ang II (10 –7 M). Brown staining in cytoplasm and nuclei represented protein expression of p-p38MAPK. Figure 4 A, A : Control group; B – F : Ang II induced for 2 minutes–30 minutes; G – H : Ena. (10 –6 M) treated for 5 and 15 minutes; I : NAC (10 –2 M, 30 minutes prior to AngII administration) for 15 min. # P

    Techniques Used: Translocation Assay, Inverted Microscopy, Immunocytochemistry, Staining, Cell Culture, Modification, Expressing

    4) Product Images from "Investigation of Immune-Regulatory Effects of Mageumsan Hot Spring via Protein Microarray In Vitro"

    Article Title: Investigation of Immune-Regulatory Effects of Mageumsan Hot Spring via Protein Microarray In Vitro

    Journal: Annals of Dermatology

    doi: 10.5021/ad.2018.30.3.322

    Cytokine protein microarray in the human keratinocyte cell cultured after 24 hours. (A) Dulbeco's Modified Eagle Medium (DMEM; Gibco-BRL, USA) media (without fetal bovine serum, osmorality 336 mOSM/kg). (B) DMEM media with Magumsan hot spring (HS) water (osmorality 350 mOSM/kg). (C) DMEM media treated with toll-like receptor 3 agonist poly (I:C) (10 µg/ml). (D) DMEM media with Magumsan HS water treated with poly (I:C) (10 µg/ml).
    Figure Legend Snippet: Cytokine protein microarray in the human keratinocyte cell cultured after 24 hours. (A) Dulbeco's Modified Eagle Medium (DMEM; Gibco-BRL, USA) media (without fetal bovine serum, osmorality 336 mOSM/kg). (B) DMEM media with Magumsan hot spring (HS) water (osmorality 350 mOSM/kg). (C) DMEM media treated with toll-like receptor 3 agonist poly (I:C) (10 µg/ml). (D) DMEM media with Magumsan HS water treated with poly (I:C) (10 µg/ml).

    Techniques Used: Microarray, Cell Culture, Modification

    5) Product Images from "Wnt2 contributes to the progression of gastric cancer by promoting cell migration and invasion"

    Article Title: Wnt2 contributes to the progression of gastric cancer by promoting cell migration and invasion

    Journal: Oncology Letters

    doi: 10.3892/ol.2018.9050

    Secreted Wnt2 promotes cell proliferation in gastric cancer cell lines. A plasmid containing Wnt2 was stably transfected into CHO cells and CM containing secreted Wnt2 was collected for further study. (A) Expression of Wnt2 in CHO-W was confirmed by western blot analysis. (B) Quantification of the western blot analysis indicated that Wnt2 protein could be detected in CHO-W and the CM of CHO-W. Effects of normal growth media (DMEM + 10% fetal bovine serum), VecCM or VecWnt2CM on the growth of gastric cancer cells (C) SGC-7901 and (D) BGC823 were compared by an MTT assay. Data are presented as the mean ± standard deviation of three independent experiments. CM, conditioned medium; CHO-W, Wnt2-transfected CHO cells; CHO-V, empty vector-transfected CHO cells; DMEM, Dulbecco's modified Eagle's medium; VecCM, control CM; VecWnt2CM, CHO-Wnt2 CM; OD, optical density.
    Figure Legend Snippet: Secreted Wnt2 promotes cell proliferation in gastric cancer cell lines. A plasmid containing Wnt2 was stably transfected into CHO cells and CM containing secreted Wnt2 was collected for further study. (A) Expression of Wnt2 in CHO-W was confirmed by western blot analysis. (B) Quantification of the western blot analysis indicated that Wnt2 protein could be detected in CHO-W and the CM of CHO-W. Effects of normal growth media (DMEM + 10% fetal bovine serum), VecCM or VecWnt2CM on the growth of gastric cancer cells (C) SGC-7901 and (D) BGC823 were compared by an MTT assay. Data are presented as the mean ± standard deviation of three independent experiments. CM, conditioned medium; CHO-W, Wnt2-transfected CHO cells; CHO-V, empty vector-transfected CHO cells; DMEM, Dulbecco's modified Eagle's medium; VecCM, control CM; VecWnt2CM, CHO-Wnt2 CM; OD, optical density.

    Techniques Used: Plasmid Preparation, Stable Transfection, Transfection, Expressing, Western Blot, MTT Assay, Standard Deviation, Modification

    Related Articles

    Cell Culture:

    Article Title: Neuropeptide Y, substance P, and human bone morphogenetic protein 2 stimulate human osteoblast osteogenic activity by enhancing gap junction intercellular communication
    Article Snippet: .. The cells were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco, USA) supplemented with 10% fetal bovine serum (Gibco), 100 U/mL penicillin and 100 μg/mL streptomycin. ..

    Article Title: A Cellulose/Laponite Interpenetrated Polymer Network (IPN) Hydrogel: Controllable Double-Network Structure with High Modulus
    Article Snippet: Cell Viability Cell culture: prior any biocompatibility experiments, Si-HPMC basic solution, and XLS solutions were autoclaved at 120 °C during 20 min with an Alphaklave 23. .. Human chondrosarcoma cell line (SW1353, ATCC, Molsheim, France) was cultured in a 5% CO2 incubator at 37 °C in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen Corp., Carlsbad, CA, US). ..

    Modification:

    Article Title: Neuropeptide Y, substance P, and human bone morphogenetic protein 2 stimulate human osteoblast osteogenic activity by enhancing gap junction intercellular communication
    Article Snippet: .. The cells were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco, USA) supplemented with 10% fetal bovine serum (Gibco), 100 U/mL penicillin and 100 μg/mL streptomycin. ..

    Article Title: Identification of Translational Activators of Glial Glutamate Transporter EAAT2 through Cell-Based High-Throughput Screening: An Approach to Prevent Excitotoxicity
    Article Snippet: PA-EAAT2 cells, a primary astrocyte line that stably expresses human EAAT2 transcripts with the 1091-nucleotides 5′ UTR driven by the cytomegalovirus (CMV) promoter, were generated in our previous study. .. PA-EAAT2 cells at passage 9 were thawed from frozen stocks and grown in Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen, Carlsbad, CA) containing 25 mM glucose, 1 mM sodium pyruvate, 19.4 μM pyridoxine hydrochloride, and 2 mM glutamine and supplemented with 10% fetal bovine serum (FBS), 700 μg/mL geneticin (Gibco, Los Angeles, CA), and 100 μg/mL penicillin-streptomycin (Sigma, St. Louis, MO). ..

    Article Title: Chemical Reactive Anchoring Lipids with Different Performance for Cell Surface Re-engineering Application
    Article Snippet: Azido–PEG4 –biotin and streptavidin-fluorescein isothiocyanate (streptavidin-FITC) were purchased from Biolegend (San Diego, CA). .. Dulbecco’s modified Eagle’s medium (DMEM), (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), and MTT reagent were purchased from Life Technologies (Grand Island, NY). .. First, monocholesteryl–PEG2000 -amine was synthesized as per a previously described method.

    Article Title: A Cellulose/Laponite Interpenetrated Polymer Network (IPN) Hydrogel: Controllable Double-Network Structure with High Modulus
    Article Snippet: Cell Viability Cell culture: prior any biocompatibility experiments, Si-HPMC basic solution, and XLS solutions were autoclaved at 120 °C during 20 min with an Alphaklave 23. .. Human chondrosarcoma cell line (SW1353, ATCC, Molsheim, France) was cultured in a 5% CO2 incubator at 37 °C in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen Corp., Carlsbad, CA, US). ..

    Article Title: The Human Papillomavirus Type 8 E6 Protein Interferes with NOTCH Activation during Keratinocyte Differentiation
    Article Snippet: Therefore, we tested if calcium treatment of iHFKs resulted in a NOTCH activity profile similar to that of differentiating skin, namely, a gradual but robust increase of signaling during differentiation ( ). .. 8E6-iHFKs and C-iHFKs were grown to confluence in standard keratinocyte serum-free media (KFSM) supplemented with bovine pituitary gland extract and epidermal growth factor (Gibco) and then switched to calcium-containing Dulbecco's modified Eagle medium (DMEM; Gibco) supplemented with 10% fetal bovine serum. .. Total RNA was harvested with an RNAeasy Plus minikit (Qiagen) at days 0, 3, 4, 5, and 6 after calcium treatment, and relative mRNA levels of HES1 and MAML1 were determined as described for .

    Article Title: Characterization of Virulent West Nile Virus Kunjin Strain, Australia, 2011
    Article Snippet: The mosquitoes were submitted live to the Medical Entomology Laboratory at Westmead Hospital (Westmead, NSW, Australia) for species identification, arbovirus isolation, and virus identification ( ). .. Cells and Viruses We propagated Vero 76 cells in Dulbecco modified minimum essential medium (DMEM; Life Technologies) supplemented with 10% fetal bovine serum (FBS). ..

    Article Title: Selenate Prevents Adipogenesis through Induction of Selenoprotein S and Attenuation of Endoplasmic Reticulum Stress
    Article Snippet: The fetal calf serum (FCS) and fetal bovine serum (FBS) were purchased from the PAA Cell Culture Company (Worcester, MA, USA). .. The Dulbecco’s Modified Eagle’s medium (DMEM) and 0.25% trypsin-EDTA were obtained from Thermo Fisher Scientific (Waltham, MA, USA). .. The rosiglitazone was purchased from Calbiochem (San Diego, CA, USA).

    Article Title: Mesenchymal Stromal Cells Engineered to Produce IGF-I by Recombinant Adenovirus Ameliorate Liver Fibrosis in Mice
    Article Snippet: Mononuclear cells were isolated using Ficoll–PaqueTM Plus density gradient (1.077 g/mL; GE Healthcare). .. Cells were plated at 4,000 cells per cm2 and incubated in Dulbecco's modified Eagle's medium low glucose (DMEM lg; Invitrogen/Life Technologies) supplemented with 10% fetal bovine serum (FBS; Gibco/Invitrogen). ..

    MTT Assay:

    Article Title: Chemical Reactive Anchoring Lipids with Different Performance for Cell Surface Re-engineering Application
    Article Snippet: Azido–PEG4 –biotin and streptavidin-fluorescein isothiocyanate (streptavidin-FITC) were purchased from Biolegend (San Diego, CA). .. Dulbecco’s modified Eagle’s medium (DMEM), (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), and MTT reagent were purchased from Life Technologies (Grand Island, NY). .. First, monocholesteryl–PEG2000 -amine was synthesized as per a previously described method.

    Incubation:

    Article Title: Mesenchymal Stromal Cells Engineered to Produce IGF-I by Recombinant Adenovirus Ameliorate Liver Fibrosis in Mice
    Article Snippet: Mononuclear cells were isolated using Ficoll–PaqueTM Plus density gradient (1.077 g/mL; GE Healthcare). .. Cells were plated at 4,000 cells per cm2 and incubated in Dulbecco's modified Eagle's medium low glucose (DMEM lg; Invitrogen/Life Technologies) supplemented with 10% fetal bovine serum (FBS; Gibco/Invitrogen). ..

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    Thermo Fisher dmem
    FasL is generated in Rac1-expressing cells after serum deprivation and it is necessary for apoptosis. (A) <t>NIH</t> 3T3 cells stably expressing Ras-1QL protein (Rac1) or control NIH 3T3 cells (Control) were incubated in <t>DMEM</t> with (+) or without (−) serum for 24 h. Then, cells were processed for FasL expression determined by RT-PCR as described in MATERIALS AND METHODS. FasL expression in shown on the left (Fas Ligand), and as control, the β-actin expression (β-Actin) was determined with the same samples in parallel reactions. Negative control refers to an RT-PCR amplification without primers. (B) Western blot analysis of FasL levels in NIH 3T3 and Rac1 cells in the presence and absence of serum. (C) Cells were incubated with FasFc (0.2 μg/ml) either in control medium or without serum for 24 h. Cell death was estimated by flow cytometry (Annexin V) and expressed as percentage of the value of Rac1 without serum for each experiment (cell death range: 22–38%). Data are the mean ± SD from three independent experiments.
    Dmem, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher dulbecco s modified eagle s medium
    (A) The expression of elastin (ELN, green) colocalized with the tenocytes as labelled by tenascin-C (TN-C, red) in normal tendon (incubated in DMEM). ELNase specifically degrades ELN but does not disrupt the fascicle, as illustrated in the TN-C staining. However, COLase caused severe diffusion of the ELN and TN-C expressions. (B) The increase in COL production during ELNase incubation is confirmed by specific immunofluorescence staining of COL. (C) Both ELNase and COLase increased the cyclooxygenase-2 (COX-2) expressions to induce the inflammation in tenocytes. Scale bar ​= ​50 ​μm. DMEM = <t>Dulbecco's</t> Modified <t>Eagle's</t> Medium.
    Dulbecco S Modified Eagle S Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Thermo Fisher mesc medium
    SIRT1 expression during iPSC formation and differentiation of ESCs and iPSCs in mouse model. (A) Temporal expression of Sirt1 mRNA on day 0 to day 20 after DOX treatment. MEF without DOX on day 20 (20-DOX) and <t>mESC</t> were included. (B) Western blotting analysis of SIRT1, OCT4 and PCNA in 2°F/1B MEF without (-DOX) and with (+DOX) DOX treatment for 15 days, serially passaged iPSC from passages 4 (P4), 5–7 (P5, P6, P7) and differentiated colonies at passage 4 (Diff-P4). (C) The relative expression levels of SIRT1 protein in MEF, <t>miPSC</t> and mESC. (D) Relative SIRT1 protein expressions in embryoid bodies collected from mESC and miPSC on days 2, 5, 8, 11, 14 and 17 after differentiation. D0 are the undifferentiated cell control. *p
    Mesc Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    FasL is generated in Rac1-expressing cells after serum deprivation and it is necessary for apoptosis. (A) NIH 3T3 cells stably expressing Ras-1QL protein (Rac1) or control NIH 3T3 cells (Control) were incubated in DMEM with (+) or without (−) serum for 24 h. Then, cells were processed for FasL expression determined by RT-PCR as described in MATERIALS AND METHODS. FasL expression in shown on the left (Fas Ligand), and as control, the β-actin expression (β-Actin) was determined with the same samples in parallel reactions. Negative control refers to an RT-PCR amplification without primers. (B) Western blot analysis of FasL levels in NIH 3T3 and Rac1 cells in the presence and absence of serum. (C) Cells were incubated with FasFc (0.2 μg/ml) either in control medium or without serum for 24 h. Cell death was estimated by flow cytometry (Annexin V) and expressed as percentage of the value of Rac1 without serum for each experiment (cell death range: 22–38%). Data are the mean ± SD from three independent experiments.

    Journal: Molecular Biology of the Cell

    Article Title: Apoptosis Induced by Rac GTPase Correlates with Induction of FasL and Ceramides Production

    doi:

    Figure Lengend Snippet: FasL is generated in Rac1-expressing cells after serum deprivation and it is necessary for apoptosis. (A) NIH 3T3 cells stably expressing Ras-1QL protein (Rac1) or control NIH 3T3 cells (Control) were incubated in DMEM with (+) or without (−) serum for 24 h. Then, cells were processed for FasL expression determined by RT-PCR as described in MATERIALS AND METHODS. FasL expression in shown on the left (Fas Ligand), and as control, the β-actin expression (β-Actin) was determined with the same samples in parallel reactions. Negative control refers to an RT-PCR amplification without primers. (B) Western blot analysis of FasL levels in NIH 3T3 and Rac1 cells in the presence and absence of serum. (C) Cells were incubated with FasFc (0.2 μg/ml) either in control medium or without serum for 24 h. Cell death was estimated by flow cytometry (Annexin V) and expressed as percentage of the value of Rac1 without serum for each experiment (cell death range: 22–38%). Data are the mean ± SD from three independent experiments.

    Article Snippet: NIH 3T3 cells were maintained in DMEM (Life Technologies, Gaithersburg, MD) supplemented with 10% newborn calf serum (NCS) under standard conditions of temperature (37°C), humidity (95°C), and carbon dioxide (5%).

    Techniques: Generated, Expressing, Stable Transfection, Incubation, Reverse Transcription Polymerase Chain Reaction, Negative Control, Amplification, Western Blot, Flow Cytometry, Cytometry

    Induction of apoptosis by exogenous addition of FasL and/or synthetic ceramides. Analysis of DNA fragmentation was performed essentially as described in MATERIALS AND METHODS. Control, NIH 3T3 cells, and NIH 3T3 overexpressing Rac1 were incubated in DMEM supplemented with 10% NCS until they reached 70% confluence. At this time, cells were incubated in DMEM with (+) or without (−) serum, and treated as follows. (A) Cells were incubated with Fas ligand (50 ng/ml) in the absence or presence of serum for 19 h. (B) Cells were incubated with C 2 ceramide (100 μM) in the presence or absence of serum for 48 h. (C) Cells were incubated with Fas ligand (50 ng/ml) plus ceramides C 2 (100 μM) in the presence or absence of 10% NCS for 24 h.

    Journal: Molecular Biology of the Cell

    Article Title: Apoptosis Induced by Rac GTPase Correlates with Induction of FasL and Ceramides Production

    doi:

    Figure Lengend Snippet: Induction of apoptosis by exogenous addition of FasL and/or synthetic ceramides. Analysis of DNA fragmentation was performed essentially as described in MATERIALS AND METHODS. Control, NIH 3T3 cells, and NIH 3T3 overexpressing Rac1 were incubated in DMEM supplemented with 10% NCS until they reached 70% confluence. At this time, cells were incubated in DMEM with (+) or without (−) serum, and treated as follows. (A) Cells were incubated with Fas ligand (50 ng/ml) in the absence or presence of serum for 19 h. (B) Cells were incubated with C 2 ceramide (100 μM) in the presence or absence of serum for 48 h. (C) Cells were incubated with Fas ligand (50 ng/ml) plus ceramides C 2 (100 μM) in the presence or absence of 10% NCS for 24 h.

    Article Snippet: NIH 3T3 cells were maintained in DMEM (Life Technologies, Gaithersburg, MD) supplemented with 10% newborn calf serum (NCS) under standard conditions of temperature (37°C), humidity (95°C), and carbon dioxide (5%).

    Techniques: Incubation

    Effect of caspase protease inhibitors on Rac-induced apoptosis. Rac1 and control NIH 3T3 cells were incubated in either DMEM supplemented with 10% NCS (+) or DMEM without serum (−). The caspase-3 inhibitor acetyl-DEVD-CHO (50 μM) (A) or the caspase-1 inhibitor acetyl-YVAD-CHO (100 μM) (B) was added and cells incubated for an additional period of 24 h. After this period, cells were analyzed for induction of apoptosis by the DNA fragmentation analysis as described in MATERIALS AND METHODS. Where indicated, DMEM supplemented with PDGF (100 nM) was used. Control, NIH 3T3 cells; Rac1, NIH 3T3 cells overexpressing the mutated Rac1-QL protein.

    Journal: Molecular Biology of the Cell

    Article Title: Apoptosis Induced by Rac GTPase Correlates with Induction of FasL and Ceramides Production

    doi:

    Figure Lengend Snippet: Effect of caspase protease inhibitors on Rac-induced apoptosis. Rac1 and control NIH 3T3 cells were incubated in either DMEM supplemented with 10% NCS (+) or DMEM without serum (−). The caspase-3 inhibitor acetyl-DEVD-CHO (50 μM) (A) or the caspase-1 inhibitor acetyl-YVAD-CHO (100 μM) (B) was added and cells incubated for an additional period of 24 h. After this period, cells were analyzed for induction of apoptosis by the DNA fragmentation analysis as described in MATERIALS AND METHODS. Where indicated, DMEM supplemented with PDGF (100 nM) was used. Control, NIH 3T3 cells; Rac1, NIH 3T3 cells overexpressing the mutated Rac1-QL protein.

    Article Snippet: NIH 3T3 cells were maintained in DMEM (Life Technologies, Gaithersburg, MD) supplemented with 10% newborn calf serum (NCS) under standard conditions of temperature (37°C), humidity (95°C), and carbon dioxide (5%).

    Techniques: Incubation

    Caspase-3 activation in apoptosis induced by Rac1. NIH 3T3 cells constitutively expressing the Rac1QL protein and cells transfected with the empty plasmid (c) were cultivated for 24 h in DMEM supplemented with 10% new born calf serum (+) or DMEM without serum (−). The cells were collected at 0, 6, 12, and 24 h after treatment and cytoplasmic extracts were tested for protease activity by using specific substrates as indicated in MATERIALS AND METHODS. Caspase-1 and caspase-3 enzyme activity is expressed as fold activation over basal levels. Data are representative of four experiments performed in triplicates, with similar results. Bars represent SD.

    Journal: Molecular Biology of the Cell

    Article Title: Apoptosis Induced by Rac GTPase Correlates with Induction of FasL and Ceramides Production

    doi:

    Figure Lengend Snippet: Caspase-3 activation in apoptosis induced by Rac1. NIH 3T3 cells constitutively expressing the Rac1QL protein and cells transfected with the empty plasmid (c) were cultivated for 24 h in DMEM supplemented with 10% new born calf serum (+) or DMEM without serum (−). The cells were collected at 0, 6, 12, and 24 h after treatment and cytoplasmic extracts were tested for protease activity by using specific substrates as indicated in MATERIALS AND METHODS. Caspase-1 and caspase-3 enzyme activity is expressed as fold activation over basal levels. Data are representative of four experiments performed in triplicates, with similar results. Bars represent SD.

    Article Snippet: NIH 3T3 cells were maintained in DMEM (Life Technologies, Gaithersburg, MD) supplemented with 10% newborn calf serum (NCS) under standard conditions of temperature (37°C), humidity (95°C), and carbon dioxide (5%).

    Techniques: Activation Assay, Expressing, Transfection, Plasmid Preparation, Activity Assay

    Rac1-induced apoptosis progress in the absence of cytochrome c accumulation in the cytosol. (Left) NIH 3T3 cells (Control) and Rac1-expressing cells (Rac1) were incubated in DMEM medium with (+) or without (−) serum for 24 h. After that, cells extracts were obtained and processed for cytochrome c analysis. Both cytosolic (Cytosol) and mitochondrial preparations (Mitochond.) were tested for the presence of cytochrome c by Western blot analysis, by using a monoclonal antibody against cytochrome c . Cyt. C indicates purified cytochrome c . The arrow indicates the position of the cytochrome c . (Right) Parallel cultures of NIH 3T3 cells (Control) and Rac1-expressing cells (Rac1) were incubated in DMEM medium with (+) or without (−) serum for 24 h cells and then analyzed for induction of apoptosis by the DNA fragmentation analysis as described in MATERIALS AND METHODS.

    Journal: Molecular Biology of the Cell

    Article Title: Apoptosis Induced by Rac GTPase Correlates with Induction of FasL and Ceramides Production

    doi:

    Figure Lengend Snippet: Rac1-induced apoptosis progress in the absence of cytochrome c accumulation in the cytosol. (Left) NIH 3T3 cells (Control) and Rac1-expressing cells (Rac1) were incubated in DMEM medium with (+) or without (−) serum for 24 h. After that, cells extracts were obtained and processed for cytochrome c analysis. Both cytosolic (Cytosol) and mitochondrial preparations (Mitochond.) were tested for the presence of cytochrome c by Western blot analysis, by using a monoclonal antibody against cytochrome c . Cyt. C indicates purified cytochrome c . The arrow indicates the position of the cytochrome c . (Right) Parallel cultures of NIH 3T3 cells (Control) and Rac1-expressing cells (Rac1) were incubated in DMEM medium with (+) or without (−) serum for 24 h cells and then analyzed for induction of apoptosis by the DNA fragmentation analysis as described in MATERIALS AND METHODS.

    Article Snippet: NIH 3T3 cells were maintained in DMEM (Life Technologies, Gaithersburg, MD) supplemented with 10% newborn calf serum (NCS) under standard conditions of temperature (37°C), humidity (95°C), and carbon dioxide (5%).

    Techniques: Expressing, Incubation, Western Blot, Purification

    Induction of apoptosis by overexpression of the human Rac protein needs mRNA and proteins synthesis. (A) NIH 3T3 cells were transfected with the appropriate plasmids carrying the human gene rac -1 activated by a Leu 61 mutation (QL mutant). Pools of transfected cells were selected for geneticin resistance and equivalent amounts of protein lysates were analyzed by Western blot for the level of expression of the protein by using a specific antibody. Control indicates extracts from cells transfected with the empty vector, pLNCX. Rac1 indicates the constitutively active pLNCX-Rac1 QL mutant. The arrow on the right indicates the position of Rac1QL protein. (B) Analysis of apoptosis by the DNA fragmentation assay after serum deprivation. Cells were incubated for 24 h in DMEM with or without serum and in presence of 50 μM cycloheximide (CHX), actinomycin D (ACT. D). Gel shows the DNA ladder after staining with ethidium bromide.

    Journal: Molecular Biology of the Cell

    Article Title: Apoptosis Induced by Rac GTPase Correlates with Induction of FasL and Ceramides Production

    doi:

    Figure Lengend Snippet: Induction of apoptosis by overexpression of the human Rac protein needs mRNA and proteins synthesis. (A) NIH 3T3 cells were transfected with the appropriate plasmids carrying the human gene rac -1 activated by a Leu 61 mutation (QL mutant). Pools of transfected cells were selected for geneticin resistance and equivalent amounts of protein lysates were analyzed by Western blot for the level of expression of the protein by using a specific antibody. Control indicates extracts from cells transfected with the empty vector, pLNCX. Rac1 indicates the constitutively active pLNCX-Rac1 QL mutant. The arrow on the right indicates the position of Rac1QL protein. (B) Analysis of apoptosis by the DNA fragmentation assay after serum deprivation. Cells were incubated for 24 h in DMEM with or without serum and in presence of 50 μM cycloheximide (CHX), actinomycin D (ACT. D). Gel shows the DNA ladder after staining with ethidium bromide.

    Article Snippet: NIH 3T3 cells were maintained in DMEM (Life Technologies, Gaithersburg, MD) supplemented with 10% newborn calf serum (NCS) under standard conditions of temperature (37°C), humidity (95°C), and carbon dioxide (5%).

    Techniques: Over Expression, Transfection, Mutagenesis, Western Blot, Expressing, Plasmid Preparation, DNA Fragmentation Assay, Incubation, Activated Clotting Time Assay, Staining

    Impact of central carbon metabolism during C . jejuni intestinal colonization. The log 2 (fold change [intestine/inoculum]) in the number of transposon insertions within C . jejuni genes encoding enzymes in the tricarboxylic acid (TCA) cycle, gluconeogenesis, and the acetate switch pathways are shown and are derived from the raw data in S3 Table . Values below −6.2 indicate mutations that led to a statistically significant colonization defect. *: denotes genes showing a limited number of insertions within the library and no insertions within the pooled of mutants recovered from the intestine. The red arrows denote that the number of insertions within the gene involved in the indicated reaction was significantly reduced within the pool of mutants recovered from the mouse intestine (relative to the inoculum). Green arrows indicate that the enzyme that catalyzes the corresponding reaction does not have an insertional mutant in our mutant library. Enzymes not encoded in the C . jejuni genome are indicated with an “X.” The inset depicts the overall 13 C-excess and relative fractions of 13 C-labeled isotopologues in free and bound glucose or galactose (as indicated) derived from C . jejuni 81–176 cell surface carbohydrates after cultivation in Dulbecco’s Modified Eagle Medium (DMEM) with [3- 13 C 1 ]Ser. The colored boxes indicate the relative contributions (%] of isotopologues with 1, 2, and 3 13 C-atoms indicated as M+1, M+2, and M+3, respectively. Numbers are the means ± standard deviation (SD) of 6 measurements (see S11 Table ). Acetyl-CoA, acetyl coenzyme A; LOS, lipooligosaccharide; OAA, oxaloacetate; PEP, phosphoenolpyruvic acid; PPP, pentose phosphate pathway.

    Journal: PLoS Biology

    Article Title: Metabolic and fitness determinants for in vitro growth and intestinal colonization of the bacterial pathogen Campylobacter jejuni

    doi: 10.1371/journal.pbio.2001390

    Figure Lengend Snippet: Impact of central carbon metabolism during C . jejuni intestinal colonization. The log 2 (fold change [intestine/inoculum]) in the number of transposon insertions within C . jejuni genes encoding enzymes in the tricarboxylic acid (TCA) cycle, gluconeogenesis, and the acetate switch pathways are shown and are derived from the raw data in S3 Table . Values below −6.2 indicate mutations that led to a statistically significant colonization defect. *: denotes genes showing a limited number of insertions within the library and no insertions within the pooled of mutants recovered from the intestine. The red arrows denote that the number of insertions within the gene involved in the indicated reaction was significantly reduced within the pool of mutants recovered from the mouse intestine (relative to the inoculum). Green arrows indicate that the enzyme that catalyzes the corresponding reaction does not have an insertional mutant in our mutant library. Enzymes not encoded in the C . jejuni genome are indicated with an “X.” The inset depicts the overall 13 C-excess and relative fractions of 13 C-labeled isotopologues in free and bound glucose or galactose (as indicated) derived from C . jejuni 81–176 cell surface carbohydrates after cultivation in Dulbecco’s Modified Eagle Medium (DMEM) with [3- 13 C 1 ]Ser. The colored boxes indicate the relative contributions (%] of isotopologues with 1, 2, and 3 13 C-atoms indicated as M+1, M+2, and M+3, respectively. Numbers are the means ± standard deviation (SD) of 6 measurements (see S11 Table ). Acetyl-CoA, acetyl coenzyme A; LOS, lipooligosaccharide; OAA, oxaloacetate; PEP, phosphoenolpyruvic acid; PPP, pentose phosphate pathway.

    Article Snippet: For growth in liquid defined minimal medium, 108 CFUs of the C . jejuni transposon mutant library were added to 4 ml of DMEM (GIBCO; catalogue number 11965) supplemented with 20 mM Asp, Gln, or Ser.

    Techniques: Derivative Assay, Mutagenesis, Labeling, Modification, Standard Deviation

    Biosynthetic capacities of C . jejuni 81–176 upon catabolism of [3- 13 C 1 ]Ser. The intermediary metabolism of C . jejuni 81–176 was investigated through isotopologue profiling with 13 C-labelled Ser. (A) Overall 13 C-excess and relative fractions of 13 C-labeled isotopologues in protein-derived amino acids gained by acidic hydrolysis of C . jejuni 81–176 cells after cultivation in Dulbecco’s Modified Eagle Medium (DMEM) with [3- 13 C 1 ]Ser as determined by gas-chromatography/mass-spectrometry (GC/MS) analysis. The colored boxes indicate the relative contributions (%) of isotopologues with 1, 2, 3, 4, 5, and 6 13 C-atoms indicated as M+1, M+2, M+3, M+4, M+5, and M+6, respectively. Values are the means ± standard deviation (SD) of 6 measurements (see S11 Table ). (B) Overview of the anabolism in C . jejuni 81–176 fueled by the catabolism of [3- 13 C 1 ]Ser. The dots illustrate the 13 C-carbon flux from [3- 13 C 1 ]Ser within the indicated molecules. Because of stereoisometry, the positioning of the 13 C-atoms within succinate and fumarate is indistinguishable; thus, the resulting possibilities of the 13 C-positions are indicated in red and orange as a 50% labeling probability for each. Green arrows display the biosynthetic pathways confirmed through isotopologue profiling. Acetyl-CoA, acetyl coenzyme A; Ala, alanine; Asp, asparagine; Glu, glutamic acid; Gly, glycine; His, histidine; Ile, isoleucine; Leu, leucine; Lys, lysine; Phe, phenylalanine; PEP, phosphoenolpyruvic acid; PPP, pentose phosphate pathway; Pro, proline; Ser, serine; TCA, tricarboxylic acid; Thr, threonine; Tyr, tyrosine; Val, valine.

    Journal: PLoS Biology

    Article Title: Metabolic and fitness determinants for in vitro growth and intestinal colonization of the bacterial pathogen Campylobacter jejuni

    doi: 10.1371/journal.pbio.2001390

    Figure Lengend Snippet: Biosynthetic capacities of C . jejuni 81–176 upon catabolism of [3- 13 C 1 ]Ser. The intermediary metabolism of C . jejuni 81–176 was investigated through isotopologue profiling with 13 C-labelled Ser. (A) Overall 13 C-excess and relative fractions of 13 C-labeled isotopologues in protein-derived amino acids gained by acidic hydrolysis of C . jejuni 81–176 cells after cultivation in Dulbecco’s Modified Eagle Medium (DMEM) with [3- 13 C 1 ]Ser as determined by gas-chromatography/mass-spectrometry (GC/MS) analysis. The colored boxes indicate the relative contributions (%) of isotopologues with 1, 2, 3, 4, 5, and 6 13 C-atoms indicated as M+1, M+2, M+3, M+4, M+5, and M+6, respectively. Values are the means ± standard deviation (SD) of 6 measurements (see S11 Table ). (B) Overview of the anabolism in C . jejuni 81–176 fueled by the catabolism of [3- 13 C 1 ]Ser. The dots illustrate the 13 C-carbon flux from [3- 13 C 1 ]Ser within the indicated molecules. Because of stereoisometry, the positioning of the 13 C-atoms within succinate and fumarate is indistinguishable; thus, the resulting possibilities of the 13 C-positions are indicated in red and orange as a 50% labeling probability for each. Green arrows display the biosynthetic pathways confirmed through isotopologue profiling. Acetyl-CoA, acetyl coenzyme A; Ala, alanine; Asp, asparagine; Glu, glutamic acid; Gly, glycine; His, histidine; Ile, isoleucine; Leu, leucine; Lys, lysine; Phe, phenylalanine; PEP, phosphoenolpyruvic acid; PPP, pentose phosphate pathway; Pro, proline; Ser, serine; TCA, tricarboxylic acid; Thr, threonine; Tyr, tyrosine; Val, valine.

    Article Snippet: For growth in liquid defined minimal medium, 108 CFUs of the C . jejuni transposon mutant library were added to 4 ml of DMEM (GIBCO; catalogue number 11965) supplemented with 20 mM Asp, Gln, or Ser.

    Techniques: Labeling, Derivative Assay, Modification, Gas Chromatography, Mass Spectrometry, Gas Chromatography-Mass Spectrometry, Standard Deviation

    C . jejuni 81–176 fitness determinants identified by insertion sequencing (INSeq) analyses. (A) Diagram of the INSeq strategy used in these studies. (B) Histogram depicting the number of genes ( y axis) that exhibited the indicated log 2 (fold change [output/input]) change ( x axis) in the numbers of transposon insertions recovered after growth on solid rich medium relative to the number of transposon insertions in the original inoculum. Areas colored with green represent genes whose number of transposon insertions showed a significant decrease after growth on solid rich medium. (C) Venn diagram showing the relationship between genes required for growth under different culture conditions identified in these studies. (D) Venn diagram depicting the relationship between genes whose inactivation led to increased growth in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with different amino acids. Twenty-two out of 34 genes whose mutation led to increased growth encode proteins associated with the flagellar motility system (see S2 Table for details). Asn, asparagine; CFU, colony-forming unit; Gln, glutamine; Ser, serine.

    Journal: PLoS Biology

    Article Title: Metabolic and fitness determinants for in vitro growth and intestinal colonization of the bacterial pathogen Campylobacter jejuni

    doi: 10.1371/journal.pbio.2001390

    Figure Lengend Snippet: C . jejuni 81–176 fitness determinants identified by insertion sequencing (INSeq) analyses. (A) Diagram of the INSeq strategy used in these studies. (B) Histogram depicting the number of genes ( y axis) that exhibited the indicated log 2 (fold change [output/input]) change ( x axis) in the numbers of transposon insertions recovered after growth on solid rich medium relative to the number of transposon insertions in the original inoculum. Areas colored with green represent genes whose number of transposon insertions showed a significant decrease after growth on solid rich medium. (C) Venn diagram showing the relationship between genes required for growth under different culture conditions identified in these studies. (D) Venn diagram depicting the relationship between genes whose inactivation led to increased growth in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with different amino acids. Twenty-two out of 34 genes whose mutation led to increased growth encode proteins associated with the flagellar motility system (see S2 Table for details). Asn, asparagine; CFU, colony-forming unit; Gln, glutamine; Ser, serine.

    Article Snippet: For growth in liquid defined minimal medium, 108 CFUs of the C . jejuni transposon mutant library were added to 4 ml of DMEM (GIBCO; catalogue number 11965) supplemented with 20 mM Asp, Gln, or Ser.

    Techniques: Sequencing, Modification, Mutagenesis

    The contribution of CO 2 metabolism to C . jejuni intestinal colonization. Illustrated are metabolic reactions in C . jejuni that utilize bicarbonate (H 2 CO 3 - ) (A) and the carbonic anhydrase CanB-catalyzed reaction that generates bicarbonate from CO 2 (B). Numbers indicate the log2 (fold change [intestine/inoculum]) in the number of insertions in the indicated genes and are derived from the raw data in S3 Table . Values below −6.2 indicate mutations that led to a statistically significant colonization defect. Green arrows indicate mutations that led to a statistically significant colonization defect. (C) Incorporation of CO 2 into amino acids after C . jejuni 81–176 cultivation in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with H 13 CO 3 - . Shown are the overall 13 C-excess (%) (upper panel) and relative fractions of 13 C-labeled isotopologues (lower panel) in protein-derived amino acids of C . jejuni 81–176 cultivated in DMEM supplemented with 44 mM 13 C-labeled hydrogen carbonate. The colored boxes indicate the relative contributions (%) of isotopologues with 1, 2, 3, 4, 5, and 6 13 C-atoms (M+1, M+2, M+3, M+4, M+5, and M+6). Numbers are the means ± standard deviation (SD) of 6 measurements (see S11 Table ). (D) Heat map for the overall 13 C-excess of labeled amino acids in C . jejuni 81–176 wild-type and the respective pycA :: Cm mutant strain after growth with [ 13 C]bicarbonate and 20 mM lactate (upper panel) or 20 mM Ser (lower panel) as carbon and energy sources. The values of the color map depict the mean of 2 biological experiments measured in triplicate (see S11 Table ). (E) Growth analysis of the C . jejuni 81–176 pycA mutant (grey column) compared to the wild type (black column) when cultivated in DMEM supplemented with 20 mM of different carbon and energy sources. Values represent the mean values ± SD of 3 independent experiments (see S12 Table ). Ala, alanine; Asp, asparagine; Glu, glutamic acid; Gly, glycine; Ile, isoleucine; Lac, lactate; Leu, leucine; Lys, lysine; Phe, phenylalanine; Pro, proline; Ser, serine; Thr, threonine; Tyr, tyrosine; Val, valine; w/o, without; WT, wild type.

    Journal: PLoS Biology

    Article Title: Metabolic and fitness determinants for in vitro growth and intestinal colonization of the bacterial pathogen Campylobacter jejuni

    doi: 10.1371/journal.pbio.2001390

    Figure Lengend Snippet: The contribution of CO 2 metabolism to C . jejuni intestinal colonization. Illustrated are metabolic reactions in C . jejuni that utilize bicarbonate (H 2 CO 3 - ) (A) and the carbonic anhydrase CanB-catalyzed reaction that generates bicarbonate from CO 2 (B). Numbers indicate the log2 (fold change [intestine/inoculum]) in the number of insertions in the indicated genes and are derived from the raw data in S3 Table . Values below −6.2 indicate mutations that led to a statistically significant colonization defect. Green arrows indicate mutations that led to a statistically significant colonization defect. (C) Incorporation of CO 2 into amino acids after C . jejuni 81–176 cultivation in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with H 13 CO 3 - . Shown are the overall 13 C-excess (%) (upper panel) and relative fractions of 13 C-labeled isotopologues (lower panel) in protein-derived amino acids of C . jejuni 81–176 cultivated in DMEM supplemented with 44 mM 13 C-labeled hydrogen carbonate. The colored boxes indicate the relative contributions (%) of isotopologues with 1, 2, 3, 4, 5, and 6 13 C-atoms (M+1, M+2, M+3, M+4, M+5, and M+6). Numbers are the means ± standard deviation (SD) of 6 measurements (see S11 Table ). (D) Heat map for the overall 13 C-excess of labeled amino acids in C . jejuni 81–176 wild-type and the respective pycA :: Cm mutant strain after growth with [ 13 C]bicarbonate and 20 mM lactate (upper panel) or 20 mM Ser (lower panel) as carbon and energy sources. The values of the color map depict the mean of 2 biological experiments measured in triplicate (see S11 Table ). (E) Growth analysis of the C . jejuni 81–176 pycA mutant (grey column) compared to the wild type (black column) when cultivated in DMEM supplemented with 20 mM of different carbon and energy sources. Values represent the mean values ± SD of 3 independent experiments (see S12 Table ). Ala, alanine; Asp, asparagine; Glu, glutamic acid; Gly, glycine; Ile, isoleucine; Lac, lactate; Leu, leucine; Lys, lysine; Phe, phenylalanine; Pro, proline; Ser, serine; Thr, threonine; Tyr, tyrosine; Val, valine; w/o, without; WT, wild type.

    Article Snippet: For growth in liquid defined minimal medium, 108 CFUs of the C . jejuni transposon mutant library were added to 4 ml of DMEM (GIBCO; catalogue number 11965) supplemented with 20 mM Asp, Gln, or Ser.

    Techniques: Derivative Assay, Modification, Labeling, Standard Deviation, Mutagenesis

    (A) The expression of elastin (ELN, green) colocalized with the tenocytes as labelled by tenascin-C (TN-C, red) in normal tendon (incubated in DMEM). ELNase specifically degrades ELN but does not disrupt the fascicle, as illustrated in the TN-C staining. However, COLase caused severe diffusion of the ELN and TN-C expressions. (B) The increase in COL production during ELNase incubation is confirmed by specific immunofluorescence staining of COL. (C) Both ELNase and COLase increased the cyclooxygenase-2 (COX-2) expressions to induce the inflammation in tenocytes. Scale bar ​= ​50 ​μm. DMEM = Dulbecco's Modified Eagle's Medium.

    Journal: Journal of Orthopaedic Translation

    Article Title: Sequential inflammation model for Achilles tendinopathy by elastin degradation with treadmill exercise

    doi: 10.1016/j.jot.2020.03.004

    Figure Lengend Snippet: (A) The expression of elastin (ELN, green) colocalized with the tenocytes as labelled by tenascin-C (TN-C, red) in normal tendon (incubated in DMEM). ELNase specifically degrades ELN but does not disrupt the fascicle, as illustrated in the TN-C staining. However, COLase caused severe diffusion of the ELN and TN-C expressions. (B) The increase in COL production during ELNase incubation is confirmed by specific immunofluorescence staining of COL. (C) Both ELNase and COLase increased the cyclooxygenase-2 (COX-2) expressions to induce the inflammation in tenocytes. Scale bar ​= ​50 ​μm. DMEM = Dulbecco's Modified Eagle's Medium.

    Article Snippet: To understand the ECM degradation in tendon structures, the fresh, isolated Achilles tendon samples were immediately incubated with ELNase (1 U/mL; Sigma-Aldrich, Inc.) or COLase (COLase type I, Cat# 17018029, 5 ​mg/mL; Invitrogen, Thermo Fisher Scientific Co.) in Dulbecco's Modified Eagle's Medium (DMEM; Invitrogen) at 37 ​°C for 12 ​h.

    Techniques: Expressing, Incubation, Staining, Diffusion-based Assay, Immunofluorescence, Modification

    SIRT1 expression during iPSC formation and differentiation of ESCs and iPSCs in mouse model. (A) Temporal expression of Sirt1 mRNA on day 0 to day 20 after DOX treatment. MEF without DOX on day 20 (20-DOX) and mESC were included. (B) Western blotting analysis of SIRT1, OCT4 and PCNA in 2°F/1B MEF without (-DOX) and with (+DOX) DOX treatment for 15 days, serially passaged iPSC from passages 4 (P4), 5–7 (P5, P6, P7) and differentiated colonies at passage 4 (Diff-P4). (C) The relative expression levels of SIRT1 protein in MEF, miPSC and mESC. (D) Relative SIRT1 protein expressions in embryoid bodies collected from mESC and miPSC on days 2, 5, 8, 11, 14 and 17 after differentiation. D0 are the undifferentiated cell control. *p

    Journal: PLoS ONE

    Article Title: Sirtuin 1 Facilitates Generation of Induced Pluripotent Stem Cells from Mouse Embryonic Fibroblasts through the miR-34a and p53 Pathways

    doi: 10.1371/journal.pone.0045633

    Figure Lengend Snippet: SIRT1 expression during iPSC formation and differentiation of ESCs and iPSCs in mouse model. (A) Temporal expression of Sirt1 mRNA on day 0 to day 20 after DOX treatment. MEF without DOX on day 20 (20-DOX) and mESC were included. (B) Western blotting analysis of SIRT1, OCT4 and PCNA in 2°F/1B MEF without (-DOX) and with (+DOX) DOX treatment for 15 days, serially passaged iPSC from passages 4 (P4), 5–7 (P5, P6, P7) and differentiated colonies at passage 4 (Diff-P4). (C) The relative expression levels of SIRT1 protein in MEF, miPSC and mESC. (D) Relative SIRT1 protein expressions in embryoid bodies collected from mESC and miPSC on days 2, 5, 8, 11, 14 and 17 after differentiation. D0 are the undifferentiated cell control. *p

    Article Snippet: L4 and miPSC were cultured in mESC medium [DMEM with high glucose, 100 units/ml penicillin and 100 µg/ml streptomycin (Gibco, Life Technologies), 0.1 mM MEM non-essential amino-acids (Gibco), sodium pyruvate (110 mg/L, Gibco), 50 µM beta-mercaptoethanol, 15% FBS (Gibco) and 1000 units/ml LIF (Millipore).

    Techniques: Expressing, Western Blot