dulbecco  (Thermo Fisher)


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  • 99
    Name:
    Dulbecco's Modified Eagle's Limiting Medium
    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
    Applications:
    Protein Biology|Protein Crosslinking|Protein Labeling & Crosslinking
    Size:
    500 mL
    Category:
    Labeling & Detection Products, Crosslinkers
    Score:
    85
    Buy from Supplier
    Name:
    Pierce 20X Modified Dulbecco's PBS Buffer
    Description:
    Thermo Scientific Pierce 20X Modified Dulbecco's PBS is a space-saving stock solution used for preparing physiological Na- and K-phosphate buffered saline (D-PBS) antibody diluents used in ELISA, Western blotting and other immunoassays. Features of 20X Modified Dulbecco's PBS: • Dulbecco's PBS—diluted 20-fold in water, the solution yields 8 mM sodium phosphate, 2 mM potassium phosphate, 0.14M NaCl, 10 mM KCl, pH 7.4 • Easy to use—no packets to open and no powder to dissolve • Increased accuracy—eliminates the possibility of powder remaining in a packet • Saves time—20X concentration eliminates time spent waiting for powder to dissolve • Saves space—storage as concentrated stock minimizes bench space needed for solutions Pierce Concentrated Buffers are ready to use without having to weigh and dissolve dry ingredients or to adjust the pH with concentrated acid or base. Simply dilute the stock solution with pure water and proceed with your experiment. The 20X Modified Dulbecco's PBS (phosphate-buffered saline) makes 8 mM sodium phosphate, 2 mM potassium phosphate, 0.14M NaCl, 10 mM KCl, pH 7.4, when diluted to 1X with water. Applications: • Used for wash buffers and antibody diluents in applications such as ELISA, Western blotting and other immunoassays Related Products BupH Modified Dulbecco's PBS Packs
    Catalog Number:
    28344
    Price:
    None
    Applications:
    Protein Biology
    Size:
    500 mL
    Category:
    Lab Reagents and Chemicals, General Buffers for Life Science, PBS (Non-Cell Culture) & TBS
    Score:
    85
    Buy from Supplier


    Structured Review

    Thermo Fisher dulbecco
    Effect of laminarin on apoptosis-related protein expression in LoVo cells. The LoVo cells were treated with low (400 μg/ml), middle (800 μg/ml) and high (1,600 μg/ml) concentrations of lamarin, as well as <t>Dulbecco’s</t> modified Eagle’s medium/F12 culture medium as a control and HCPT as a negative control. DR, death receptor; TRAIL, TNF-related apoptosis-inducing ligand; FADD, FAS-associated protein with death domain; Casp, caspase; HCPT, hydroxycamptothecin.
    Thermo Scientific Pierce 20X Modified Dulbecco's PBS is a space-saving stock solution used for preparing physiological Na- and K-phosphate buffered saline (D-PBS) antibody diluents used in ELISA, Western blotting and other immunoassays. Features of 20X Modified Dulbecco's PBS: • Dulbecco's PBS—diluted 20-fold in water, the solution yields 8 mM sodium phosphate, 2 mM potassium phosphate, 0.14M NaCl, 10 mM KCl, pH 7.4 • Easy to use—no packets to open and no powder to dissolve • Increased accuracy—eliminates the possibility of powder remaining in a packet • Saves time—20X concentration eliminates time spent waiting for powder to dissolve • Saves space—storage as concentrated stock minimizes bench space needed for solutions Pierce Concentrated Buffers are ready to use without having to weigh and dissolve dry ingredients or to adjust the pH with concentrated acid or base. Simply dilute the stock solution with pure water and proceed with your experiment. The 20X Modified Dulbecco's PBS (phosphate-buffered saline) makes 8 mM sodium phosphate, 2 mM potassium phosphate, 0.14M NaCl, 10 mM KCl, pH 7.4, when diluted to 1X with water. Applications: • Used for wash buffers and antibody diluents in applications such as ELISA, Western blotting and other immunoassays Related Products BupH Modified Dulbecco's PBS Packs
    https://www.bioz.com/result/dulbecco/product/Thermo Fisher
    Average 99 stars, based on 28 article reviews
    Price from $9.99 to $1999.99
    dulbecco - by Bioz Stars, 2020-01
    99/100 stars

    Images

    1) Product Images from "Laminarin-induced apoptosis in human colon cancer LoVo cells"

    Article Title: Laminarin-induced apoptosis in human colon cancer LoVo cells

    Journal: Oncology Letters

    doi: 10.3892/ol.2014.1952

    Effect of laminarin on apoptosis-related protein expression in LoVo cells. The LoVo cells were treated with low (400 μg/ml), middle (800 μg/ml) and high (1,600 μg/ml) concentrations of lamarin, as well as Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and HCPT as a negative control. DR, death receptor; TRAIL, TNF-related apoptosis-inducing ligand; FADD, FAS-associated protein with death domain; Casp, caspase; HCPT, hydroxycamptothecin.
    Figure Legend Snippet: Effect of laminarin on apoptosis-related protein expression in LoVo cells. The LoVo cells were treated with low (400 μg/ml), middle (800 μg/ml) and high (1,600 μg/ml) concentrations of lamarin, as well as Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and HCPT as a negative control. DR, death receptor; TRAIL, TNF-related apoptosis-inducing ligand; FADD, FAS-associated protein with death domain; Casp, caspase; HCPT, hydroxycamptothecin.

    Techniques Used: Expressing, Modification, Negative Control

    Effect of laminarin on Bid and tBid expression in LoVo cells. The LoVo cells were treated with low (400 μg/ml), middle (800 μg/ml) and high (1,600 μg/ml) concentrations of lamarin, as well as Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and HCPT as a negative control. HCPT, hydroxycamptothecin.
    Figure Legend Snippet: Effect of laminarin on Bid and tBid expression in LoVo cells. The LoVo cells were treated with low (400 μg/ml), middle (800 μg/ml) and high (1,600 μg/ml) concentrations of lamarin, as well as Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and HCPT as a negative control. HCPT, hydroxycamptothecin.

    Techniques Used: Expressing, Modification, Negative Control

    Effect of laminarin on LoVo cell morphology. LoVo cells were treated with (A) Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and (B) hydroxycamptothecin (5 μg/ml) as a negative control, as well as (C) low (400 μg/ml), (D) middle (800 μg/ml) and (E) high (1600 μg/ml) concentrations of laminarin. Arrows indicate apoptotic bodies.
    Figure Legend Snippet: Effect of laminarin on LoVo cell morphology. LoVo cells were treated with (A) Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and (B) hydroxycamptothecin (5 μg/ml) as a negative control, as well as (C) low (400 μg/ml), (D) middle (800 μg/ml) and (E) high (1600 μg/ml) concentrations of laminarin. Arrows indicate apoptotic bodies.

    Techniques Used: Modification, Negative Control

    2) Product Images from "Correlation of spontaneous adipocyte generation with osteogenic differentiation of porcine skin-derived stem cells"

    Article Title: Correlation of spontaneous adipocyte generation with osteogenic differentiation of porcine skin-derived stem cells

    Journal: Journal of Veterinary Science

    doi: 10.4142/jvs.2019.20.1.16

    Analysis of osteogenic differentiation in porcine skin-derived stem cells (pSSCs). (A) Morphological images of osteogenic-induced pSSCs from four individual cell lines (pSSCs-I, -II, -III, and -IV). After induction for 24 days, spontaneous lipid droplets generated during osteogenic differentiation of pSSCs were stained by Oil red O (ORO; red, positive cells). Osteogenesis was assessed by von Kossa staining for mineralization in induced cells. Control cells (Non-indu) were cultured in Dulbecco's modified Eagle's medium + 10% fetal bovine serum for the same duration. Scale bars = 100 µm (A). (B–E) Quantitative data of lipid droplet formation and osteogenic differentiation potentials from pSSCs-I, -II, -III, and -IV. Levels of lipid droplets and calcium contents in osteogenic-induced pSSCs after 24 days of osteogenic differentiation were determined by ORO and calcium deposit results, respectively. (B and C) Quantitative data based on ORO staining. Data in panel C in Fig. 1 are represented as fold-change from non-induced control cells. (D and E) Quantitative data of calcium contents. Data in panel E in Fig. 1 are represented as fold-change from non-induced control cells. OD, optical density; Non-indu, non-induction; Osteo, osteogenic induction. * Significantly higher than non-induction control ( p < 0.05). a–c Values with different letters differ significantly ( p < 0.05).
    Figure Legend Snippet: Analysis of osteogenic differentiation in porcine skin-derived stem cells (pSSCs). (A) Morphological images of osteogenic-induced pSSCs from four individual cell lines (pSSCs-I, -II, -III, and -IV). After induction for 24 days, spontaneous lipid droplets generated during osteogenic differentiation of pSSCs were stained by Oil red O (ORO; red, positive cells). Osteogenesis was assessed by von Kossa staining for mineralization in induced cells. Control cells (Non-indu) were cultured in Dulbecco's modified Eagle's medium + 10% fetal bovine serum for the same duration. Scale bars = 100 µm (A). (B–E) Quantitative data of lipid droplet formation and osteogenic differentiation potentials from pSSCs-I, -II, -III, and -IV. Levels of lipid droplets and calcium contents in osteogenic-induced pSSCs after 24 days of osteogenic differentiation were determined by ORO and calcium deposit results, respectively. (B and C) Quantitative data based on ORO staining. Data in panel C in Fig. 1 are represented as fold-change from non-induced control cells. (D and E) Quantitative data of calcium contents. Data in panel E in Fig. 1 are represented as fold-change from non-induced control cells. OD, optical density; Non-indu, non-induction; Osteo, osteogenic induction. * Significantly higher than non-induction control ( p < 0.05). a–c Values with different letters differ significantly ( p < 0.05).

    Techniques Used: Derivative Assay, Generated, Staining, Cell Culture, Modification

    3) Product Images from "Effects of bradykinin on TGF-β1-induced epithelial-mesenchymal transition in ARPE-19 cells"

    Article Title: Effects of bradykinin on TGF-β1-induced epithelial-mesenchymal transition in ARPE-19 cells

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2018.8556

    BK inhibits TGF-β1-induced epithelial-mesenchymal transitionin ARPE-19 cells, and a B2R inhibitor attenuates the role of BK. ARPE-19 cells were treated with BK, TGF-β1, BK plus TGF-β1, TGF-β1 plus HOE140, BK plus TGF-β1 plus HOE140, and Dulbecco's modified Eagle's medium for 48 h prior todetection. (A) Morphological alterations were detected using an inverted phase-contrast microscope (magnification, ×100). (B) Western blot analysis detected the expression levels of E-cadherin, α-SMA and vimentin in each group. (C) The protein expression of E-cadherin, α-SMA and vimentin was identified by immunofluorescence staining (magnification, ×200). (D) A wound healing assay tested the cell migration of a scratched edge. (E) Cells that had migrated through the Transwell chamber filter over 18 h were stained with crystal violet, and imaged at ×50 magnification. (F) The number of migrated ARPE-19 cells in each group. **P < 0.001 and *P < 0.05. TGF-β1, transforming growth factor-β1; BK, bradykinin; α-SMA, α-smooth muscle actin.
    Figure Legend Snippet: BK inhibits TGF-β1-induced epithelial-mesenchymal transitionin ARPE-19 cells, and a B2R inhibitor attenuates the role of BK. ARPE-19 cells were treated with BK, TGF-β1, BK plus TGF-β1, TGF-β1 plus HOE140, BK plus TGF-β1 plus HOE140, and Dulbecco's modified Eagle's medium for 48 h prior todetection. (A) Morphological alterations were detected using an inverted phase-contrast microscope (magnification, ×100). (B) Western blot analysis detected the expression levels of E-cadherin, α-SMA and vimentin in each group. (C) The protein expression of E-cadherin, α-SMA and vimentin was identified by immunofluorescence staining (magnification, ×200). (D) A wound healing assay tested the cell migration of a scratched edge. (E) Cells that had migrated through the Transwell chamber filter over 18 h were stained with crystal violet, and imaged at ×50 magnification. (F) The number of migrated ARPE-19 cells in each group. **P < 0.001 and *P < 0.05. TGF-β1, transforming growth factor-β1; BK, bradykinin; α-SMA, α-smooth muscle actin.

    Techniques Used: Modification, Microscopy, Western Blot, Expressing, Immunofluorescence, Staining, Wound Healing Assay, Migration

    BK inhibits TGF-β1-induced epithelial-mesenchymal transition via the TGF/Smad signaling pathway. ARPE-19 cells were treated with BK, TGF-β1, BK plus TGF-β1, TGF-β1 plus HOE140, BK plus TGF-β1 plus HOE140 and Dulbecco's modified Eagle's medium for 48 h before detection. (A) Protein expression of pSmad3 and Smad7 was identified by western blot analysis. (B) Reverse transcription-quantitative polymerase chain reaction analysis and (C) immunofluorescence staining (magnification, ×200). **P < 0.001 and *P < 0.05. TGF-β1, transforming growth factor-β1; BK, bradykinin; pSmad, phosphorylated mothers against decapentaplegic homolog.
    Figure Legend Snippet: BK inhibits TGF-β1-induced epithelial-mesenchymal transition via the TGF/Smad signaling pathway. ARPE-19 cells were treated with BK, TGF-β1, BK plus TGF-β1, TGF-β1 plus HOE140, BK plus TGF-β1 plus HOE140 and Dulbecco's modified Eagle's medium for 48 h before detection. (A) Protein expression of pSmad3 and Smad7 was identified by western blot analysis. (B) Reverse transcription-quantitative polymerase chain reaction analysis and (C) immunofluorescence staining (magnification, ×200). **P < 0.001 and *P < 0.05. TGF-β1, transforming growth factor-β1; BK, bradykinin; pSmad, phosphorylated mothers against decapentaplegic homolog.

    Techniques Used: Modification, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining

    4) Product Images from "Interleukin-4 and melatonin ameliorate high glucose and interleukin-1? stimulated inflammatory reaction in human retinal endothelial cells and retinal pigment epithelial cells"

    Article Title: Interleukin-4 and melatonin ameliorate high glucose and interleukin-1? stimulated inflammatory reaction in human retinal endothelial cells and retinal pigment epithelial cells

    Journal: Molecular Vision

    doi:

    Interleukin-4 (IL-4) and melatonin downregulated the expression of vascular endothelial growth factor (VEGF). Human retinal endothelial cells (RECs) and retinal pigment epithelial (RPE) cells were cultured to a state of subconfluency and then maintained in human endothelial-serum free medium (HE-SFM) or Dulbecco’s Modified Eagle Medium (DMEM) that contained 1% serum for 24 h for synchronization. The cells were then exposed to D-glucose (30 mM; 48 h incubation) or interleukin-1β (IL-1β; 10 ng/ml; 24 h incubation) in the presence or absence of IL-4 (40 ng/ml) or melatonin (100 μM). Total RNA was extracted, and the supernatants were harvested. VEGF expression was analyzed using quantitative real-time PCR (qPCR) and enzyme-linked immunosorbent assay (ELISA), respectively. The data are expressed as the mean±standard deviation (SD; n = 4; *p
    Figure Legend Snippet: Interleukin-4 (IL-4) and melatonin downregulated the expression of vascular endothelial growth factor (VEGF). Human retinal endothelial cells (RECs) and retinal pigment epithelial (RPE) cells were cultured to a state of subconfluency and then maintained in human endothelial-serum free medium (HE-SFM) or Dulbecco’s Modified Eagle Medium (DMEM) that contained 1% serum for 24 h for synchronization. The cells were then exposed to D-glucose (30 mM; 48 h incubation) or interleukin-1β (IL-1β; 10 ng/ml; 24 h incubation) in the presence or absence of IL-4 (40 ng/ml) or melatonin (100 μM). Total RNA was extracted, and the supernatants were harvested. VEGF expression was analyzed using quantitative real-time PCR (qPCR) and enzyme-linked immunosorbent assay (ELISA), respectively. The data are expressed as the mean±standard deviation (SD; n = 4; *p

    Techniques Used: Expressing, Cell Culture, Modification, Incubation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Standard Deviation

    5) Product Images from "Sonic hedgehog (SHH) signaling improves the angiogenic potential of Wharton’s jelly-derived mesenchymal stem cells (WJ-MSC) "

    Article Title: Sonic hedgehog (SHH) signaling improves the angiogenic potential of Wharton’s jelly-derived mesenchymal stem cells (WJ-MSC)

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-017-0653-8

    SHH pathway is active in WJ-MSC in vivo and enhances their pro-angiogenic properties. In vitro pretreated Wharton’s jelly-derived mesenchymal stem cells ( WJ-MSC ) (+ N-Shh or Cyc) were applied on top of the chorioallantoic membrane ( CAM ) of chicken embryos. Treatment was repeated after 48 h in order to maintain the effect on SHH pathway modulation in WJ-MSC. The angiogenic response was evaluated after 96 h. Recombinant fibroblast growth factor 2 ( FGF2 ), a potent angiogenic stimulator, was used as a positive control ( a ), and ( b ) Dulbecco’s modified Eagle’s medium ( DMEM ) was used as negative control (WJ-MSC vehicle). c WJ-MSC seeded in Integra Matrix ( IM ). WJ-MSC seeded in Integra Matrix plus N-Shh ( d ) and Cyc ( e ). f Assay quantification. * P
    Figure Legend Snippet: SHH pathway is active in WJ-MSC in vivo and enhances their pro-angiogenic properties. In vitro pretreated Wharton’s jelly-derived mesenchymal stem cells ( WJ-MSC ) (+ N-Shh or Cyc) were applied on top of the chorioallantoic membrane ( CAM ) of chicken embryos. Treatment was repeated after 48 h in order to maintain the effect on SHH pathway modulation in WJ-MSC. The angiogenic response was evaluated after 96 h. Recombinant fibroblast growth factor 2 ( FGF2 ), a potent angiogenic stimulator, was used as a positive control ( a ), and ( b ) Dulbecco’s modified Eagle’s medium ( DMEM ) was used as negative control (WJ-MSC vehicle). c WJ-MSC seeded in Integra Matrix ( IM ). WJ-MSC seeded in Integra Matrix plus N-Shh ( d ) and Cyc ( e ). f Assay quantification. * P

    Techniques Used: In Vivo, In Vitro, Derivative Assay, Chick Chorioallantoic Membrane Assay, Recombinant, Positive Control, Modification, Negative Control

    6) Product Images from "Cytotoxic and Antiproliferative Effect of Tepary Bean Lectins on C33-A, MCF-7, SKNSH, and SW480 Cell Lines"

    Article Title: Cytotoxic and Antiproliferative Effect of Tepary Bean Lectins on C33-A, MCF-7, SKNSH, and SW480 Cell Lines

    Journal: Molecules

    doi: 10.3390/molecules19079610

    Effect of lectin on cell proliferation after 24 h during which lectins were removed from the cells, employing the 3 [H]-thymidine assay. The cells were previously incubated for 24 h with lectin solution; then, the solution was removed and the cells were incubated for only 24 h with the cell culture solutions [Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS)]. All values are expressed as mean ± SEM of three independent experiments.
    Figure Legend Snippet: Effect of lectin on cell proliferation after 24 h during which lectins were removed from the cells, employing the 3 [H]-thymidine assay. The cells were previously incubated for 24 h with lectin solution; then, the solution was removed and the cells were incubated for only 24 h with the cell culture solutions [Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS)]. All values are expressed as mean ± SEM of three independent experiments.

    Techniques Used: Incubation, Cell Culture, Modification

    Cytotoxicity of lectin after 48 h, showing that the lectins were removed from the cells, employing the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell assay. The cells were previously incubated for 24 h with the lectin solution. Then, the solution was removed and the cells were incubated for only 48 h with the cell culture solution [Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS)]. All values are expressed as mean ± SEM of three independent experiments.
    Figure Legend Snippet: Cytotoxicity of lectin after 48 h, showing that the lectins were removed from the cells, employing the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell assay. The cells were previously incubated for 24 h with the lectin solution. Then, the solution was removed and the cells were incubated for only 48 h with the cell culture solution [Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS)]. All values are expressed as mean ± SEM of three independent experiments.

    Techniques Used: MTT Assay, Incubation, Cell Culture, Modification

    Cytotoxicity of lectin after 24 h, during which lectins were removed from the cells employing the 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell assay. The cells were previously incubated for 24 h with the lectin solution; then, the solutions were removed and the cells were incubated for 24 h only with cell culture solutions [Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS)]. All values are expressed as mean ± SEM of three independent experiments.
    Figure Legend Snippet: Cytotoxicity of lectin after 24 h, during which lectins were removed from the cells employing the 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell assay. The cells were previously incubated for 24 h with the lectin solution; then, the solutions were removed and the cells were incubated for 24 h only with cell culture solutions [Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS)]. All values are expressed as mean ± SEM of three independent experiments.

    Techniques Used: MTT Assay, Incubation, Cell Culture, Modification

    Effect of lectin on cell proliferation 48 h after the lectins were removed from the cells, employing the 3 [H]-thymidine assay. Cells were previously incubated for 24 h with the lectin solutions; then, the solutions were removed and the cells were incubated for only 48 h with the culture cell solutions [Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS)]. All values are expressed as mean ± SEM of three independent experiments.
    Figure Legend Snippet: Effect of lectin on cell proliferation 48 h after the lectins were removed from the cells, employing the 3 [H]-thymidine assay. Cells were previously incubated for 24 h with the lectin solutions; then, the solutions were removed and the cells were incubated for only 48 h with the culture cell solutions [Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS)]. All values are expressed as mean ± SEM of three independent experiments.

    Techniques Used: Incubation, Modification

    7) Product Images from "Copper as an alternative antimicrobial coating for implants - An in vitro study"

    Article Title: Copper as an alternative antimicrobial coating for implants - An in vitro study

    Journal: World Journal of Transplantation

    doi: 10.5500/wjt.v7.i3.193

    Copper release in Dulbecco’s modified Eagle medium. A high amount of copper is released from the TiCuN layer after incubation in DMEM for 24 h. The copper release is reduced on TiCuN + BONIT ® due to the BONIT ® layer. A complete exchange of the medium and seeding with MG-63 cells for another 24 h reveals significantly reduced copper release from TiCuN. The amount is equalized to the level on TiCuN + BONIT ® ( n = 3, mean value ± SD, t -test, b P
    Figure Legend Snippet: Copper release in Dulbecco’s modified Eagle medium. A high amount of copper is released from the TiCuN layer after incubation in DMEM for 24 h. The copper release is reduced on TiCuN + BONIT ® due to the BONIT ® layer. A complete exchange of the medium and seeding with MG-63 cells for another 24 h reveals significantly reduced copper release from TiCuN. The amount is equalized to the level on TiCuN + BONIT ® ( n = 3, mean value ± SD, t -test, b P

    Techniques Used: Modification, Incubation

    Scanning electron microscopy images of MG-63 osteoblasts on the pre-incubated surfaces. Samples were pre-incubated in DMEM for 24 h. After a complete exchange of medium, cells were seeded onto the surface and cultivated for another 24 h. Cells spread well on TPS and TiCuN surfaces but seem to be smaller on TiCuN + BONIT ® (magnification × 1000). TPS: Titanium plasma spray; TiCuN: Titanium-copper-nitride; DMEM: Dulbecco’s modified Eagle medium.
    Figure Legend Snippet: Scanning electron microscopy images of MG-63 osteoblasts on the pre-incubated surfaces. Samples were pre-incubated in DMEM for 24 h. After a complete exchange of medium, cells were seeded onto the surface and cultivated for another 24 h. Cells spread well on TPS and TiCuN surfaces but seem to be smaller on TiCuN + BONIT ® (magnification × 1000). TPS: Titanium plasma spray; TiCuN: Titanium-copper-nitride; DMEM: Dulbecco’s modified Eagle medium.

    Techniques Used: Electron Microscopy, Incubation, Modification

    8) Product Images from "Polygonum cuspidatum and Its Active Components Inhibit Replication of the Influenza Virus through Toll-Like Receptor 9-Induced Interferon Beta Expression"

    Article Title: Polygonum cuspidatum and Its Active Components Inhibit Replication of the Influenza Virus through Toll-Like Receptor 9-Induced Interferon Beta Expression

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0117602

    Polygonum cuspidatum (PC), resveratrol, and emodin reduced plaque numbers in a plaque reduction assay and inhibited the accumulation of hemagglutinin and neuraminidase in A549 cells. (A) Polygonum cuspidatum (PC), resveratrol, and emodin reduced plaque numbers in a plaque reduction assay. A representative plaque reduction assay is shown. Madin-Darby canine kidney (MDCK) cells were seeded into 6-well plates (1 × 10 6 cells/well) and infected with H1N1 influenza virus (100 PFU/well). The cells were treated with PC or its active components in Dulbecco’s modified Eagle medium containing 0.3% agarose. After 72 h, the plaques were determined by staining with 0.1% crystal violet. Well 1: virus control; well 2: virus + PC (1.47 mg/mL); well 3: virus + 25 μM resveratrol; well 4: mock; well 5: virus + PC (0.294 mg/mL); well 6: virus + 25 μM emodin. (B) PC, resveratrol, and emodin reduced plaque numbers in a plaque reduction assay. Results are the mean ± standard deviation of three independent experiments performed in triplicate. Asterisks indicate the calculated p values for paired comparisons between 1 and 2, 3, 5, and 6 are
    Figure Legend Snippet: Polygonum cuspidatum (PC), resveratrol, and emodin reduced plaque numbers in a plaque reduction assay and inhibited the accumulation of hemagglutinin and neuraminidase in A549 cells. (A) Polygonum cuspidatum (PC), resveratrol, and emodin reduced plaque numbers in a plaque reduction assay. A representative plaque reduction assay is shown. Madin-Darby canine kidney (MDCK) cells were seeded into 6-well plates (1 × 10 6 cells/well) and infected with H1N1 influenza virus (100 PFU/well). The cells were treated with PC or its active components in Dulbecco’s modified Eagle medium containing 0.3% agarose. After 72 h, the plaques were determined by staining with 0.1% crystal violet. Well 1: virus control; well 2: virus + PC (1.47 mg/mL); well 3: virus + 25 μM resveratrol; well 4: mock; well 5: virus + PC (0.294 mg/mL); well 6: virus + 25 μM emodin. (B) PC, resveratrol, and emodin reduced plaque numbers in a plaque reduction assay. Results are the mean ± standard deviation of three independent experiments performed in triplicate. Asterisks indicate the calculated p values for paired comparisons between 1 and 2, 3, 5, and 6 are

    Techniques Used: Infection, Modification, Staining, Standard Deviation

    9) Product Images from "MMP‐2 and MMP‐14 Silencing Inhibits VEGFR2 Cleavage and Induces the Differentiation of Porcine Adipose‐Derived Mesenchymal Stem Cells to Endothelial Cells"

    Article Title: MMP‐2 and MMP‐14 Silencing Inhibits VEGFR2 Cleavage and Induces the Differentiation of Porcine Adipose‐Derived Mesenchymal Stem Cells to Endothelial Cells

    Journal: Stem Cells Translational Medicine

    doi: 10.1002/sctm.16-0329

    (I) : Characterization of adipose‐derived mesenchymal stem cells (AMSCs). (A) AMSCs showing fibroblastoid morphology in culture; immunostaining of MSCs showing negative staining for CD14 (B), CD45 (C), and positive staining for CD44 (D), CD90 (E), and CD105 (F). Cell nuclei were stained with DAPI. (II) : Immunophenotyping of AMSCs. Flow cytometry data showing no expression for CD14 (A), and CD45 (B), and high expression for the MSC markers CD44 (C), and CD90 (D). The blue peaks show the profile of the isotype control. Flow cytometry was done on a FACS Aria Flow Cytometry System (BD Biosciences). (III) : Multilineage differentiation of AMSCs in vitro. (A) AMSCs in culture with Dulbecco's modified Eagle medium before stimulation to differentiate into any of the lineages. (B) Alcian blue staining of AMSCs culture after 20 days of stimulation with chondrogenic differentiation medium. (C) Alizarin red S staining of AMSCs culture after 15 days of stimulation with osteogenic differentiation medium. (D) Oil red O staining of AMSCs culture after 20 days of stimulation with adipogenic differentiation medium. Abbreviation: CD, cluster of differentiation.
    Figure Legend Snippet: (I) : Characterization of adipose‐derived mesenchymal stem cells (AMSCs). (A) AMSCs showing fibroblastoid morphology in culture; immunostaining of MSCs showing negative staining for CD14 (B), CD45 (C), and positive staining for CD44 (D), CD90 (E), and CD105 (F). Cell nuclei were stained with DAPI. (II) : Immunophenotyping of AMSCs. Flow cytometry data showing no expression for CD14 (A), and CD45 (B), and high expression for the MSC markers CD44 (C), and CD90 (D). The blue peaks show the profile of the isotype control. Flow cytometry was done on a FACS Aria Flow Cytometry System (BD Biosciences). (III) : Multilineage differentiation of AMSCs in vitro. (A) AMSCs in culture with Dulbecco's modified Eagle medium before stimulation to differentiate into any of the lineages. (B) Alcian blue staining of AMSCs culture after 20 days of stimulation with chondrogenic differentiation medium. (C) Alizarin red S staining of AMSCs culture after 15 days of stimulation with osteogenic differentiation medium. (D) Oil red O staining of AMSCs culture after 20 days of stimulation with adipogenic differentiation medium. Abbreviation: CD, cluster of differentiation.

    Techniques Used: Derivative Assay, Immunostaining, Negative Staining, Staining, Flow Cytometry, Cytometry, Expressing, FACS, In Vitro, Modification

    Reverse transcriptase‐polymerase chain reaction for MMPs and TIMPs mRNA expression during 10 days of stimulation with endothelial growth medium (EGM). The graphs showing mRNA expression of (A) MMP‐1, (B) MMP‐2, (C) MMP‐3, (D) MMP‐7, (E) MMP‐9, (F) MMP‐10, (G) MMP‐11, (H) MMP‐14 (MT1‐MMP), (I) TIMP‐1, and (J) TIMP‐2 in relevance to control (adipose‐derived mesenchymal stem cells with Dulbecco's modified Eagle medium) at three time points during stimulation with EGM for endothelial differentiation (*, p
    Figure Legend Snippet: Reverse transcriptase‐polymerase chain reaction for MMPs and TIMPs mRNA expression during 10 days of stimulation with endothelial growth medium (EGM). The graphs showing mRNA expression of (A) MMP‐1, (B) MMP‐2, (C) MMP‐3, (D) MMP‐7, (E) MMP‐9, (F) MMP‐10, (G) MMP‐11, (H) MMP‐14 (MT1‐MMP), (I) TIMP‐1, and (J) TIMP‐2 in relevance to control (adipose‐derived mesenchymal stem cells with Dulbecco's modified Eagle medium) at three time points during stimulation with EGM for endothelial differentiation (*, p

    Techniques Used: Polymerase Chain Reaction, Expressing, Derivative Assay, Modification

    10) Product Images from "Identification of Translational Activators of Glial Glutamate Transporter EAAT2 through Cell-Based High-Throughput Screening: An Approach to Prevent Excitotoxicity"

    Article Title: Identification of Translational Activators of Glial Glutamate Transporter EAAT2 through Cell-Based High-Throughput Screening: An Approach to Prevent Excitotoxicity

    Journal:

    doi: 10.1177/1087057110370998

    Validation of the enzyme-linked immunosorbent assay (ELISA). PA-EAAT2 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) and treated with retinoic acid (1.5 μg/mL) for 72 h and then subjected to ( A ) ELISA, ( B ) Western
    Figure Legend Snippet: Validation of the enzyme-linked immunosorbent assay (ELISA). PA-EAAT2 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) and treated with retinoic acid (1.5 μg/mL) for 72 h and then subjected to ( A ) ELISA, ( B ) Western

    Techniques Used: Enzyme-linked Immunosorbent Assay, Cell Culture, Modification, Western Blot

    Confirmation of the hits. PA-EAAT2 cells were treated with indicated concentrations of compound 9 in Dulbecco’s modified Eagle’s medium (DMEM) for 72 h and then harvested ( A ) for determining EAAT2 protein levels by Western blotting (equal
    Figure Legend Snippet: Confirmation of the hits. PA-EAAT2 cells were treated with indicated concentrations of compound 9 in Dulbecco’s modified Eagle’s medium (DMEM) for 72 h and then harvested ( A ) for determining EAAT2 protein levels by Western blotting (equal

    Techniques Used: Modification, Western Blot

    11) Product Images from "Antitumor effects of metformin via indirect inhibition of protein phosphatase 2A in patients with endometrial cancer"

    Article Title: Antitumor effects of metformin via indirect inhibition of protein phosphatase 2A in patients with endometrial cancer

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0192759

    Effects of metformin on PPP2R4 mRNA levels in vitro . HEC 265 and HEC 1B were seeded in 6-well plates (50,000 cells/well) in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum. At 80% confluency, the cells were treated with increasing doses (0.1–5 mM) of metformin for 24 h. There were no significant changes in the expression of PPP2R4 compared with the control at any concentration of metformin in either endometrial cancer cell line. The results are presented as the mean ± standard deviation for three independent experiments.
    Figure Legend Snippet: Effects of metformin on PPP2R4 mRNA levels in vitro . HEC 265 and HEC 1B were seeded in 6-well plates (50,000 cells/well) in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum. At 80% confluency, the cells were treated with increasing doses (0.1–5 mM) of metformin for 24 h. There were no significant changes in the expression of PPP2R4 compared with the control at any concentration of metformin in either endometrial cancer cell line. The results are presented as the mean ± standard deviation for three independent experiments.

    Techniques Used: In Vitro, Modification, Expressing, Concentration Assay, Standard Deviation

    12) Product Images from "Aptamer-targeted Antigen Delivery"

    Article Title: Aptamer-targeted Antigen Delivery

    Journal: Molecular Therapy

    doi: 10.1038/mt.2014.51

    Min.2 is functional for crosspresentation in vitro . ( a ) Proliferation of carboxyfluorescein succinimidyl ester (CFSE)-labeled OT-I cells following incubation of monovalent aptamer:OVA conjugates or controls with primary dendritic cells (DCs). CD11c + DCs were isolated from the spleens of C57BL/6 mice. After isolation, 4 × 10 5 DCs were incubated with pIC plus lipopolysaccharide for 4 hours (except Dulbecco's phosphate-buffered saline labeled), followed by overnight incubation with 10 nmol/l conjugate or controls for 16–20 hours at 37 °C. The next day, a single-cell suspension of 1 × 10 5 CFSE-labeled OT-I cells was added, and the cells were incubated for an additional 3 days at 37 °C, washed with flow cytometry buffer, stained with 4′,6-diamidino-2-phenylindole and antibodies against CD8α and TCR-β, and analyzed by flow cytometry. See Materials and Methods for full procedure. ( b–d ) Intracellular cytokine staining of OT-I cells in ( a ). Cells were treated with 10 μg/ml brefeldin A, incubated for 5 hours at 37 °C, stained with antibodies against CD25, interleukin (IL)-2, and interferon (IFN)-γ, and analyzed by flow cytometry. OT-I responses following incubation with min.2:OVA versus cntrl.36:OVA and NLDC145:OVA versus IgG2a:OVA were compared by student's t -test (**** P
    Figure Legend Snippet: Min.2 is functional for crosspresentation in vitro . ( a ) Proliferation of carboxyfluorescein succinimidyl ester (CFSE)-labeled OT-I cells following incubation of monovalent aptamer:OVA conjugates or controls with primary dendritic cells (DCs). CD11c + DCs were isolated from the spleens of C57BL/6 mice. After isolation, 4 × 10 5 DCs were incubated with pIC plus lipopolysaccharide for 4 hours (except Dulbecco's phosphate-buffered saline labeled), followed by overnight incubation with 10 nmol/l conjugate or controls for 16–20 hours at 37 °C. The next day, a single-cell suspension of 1 × 10 5 CFSE-labeled OT-I cells was added, and the cells were incubated for an additional 3 days at 37 °C, washed with flow cytometry buffer, stained with 4′,6-diamidino-2-phenylindole and antibodies against CD8α and TCR-β, and analyzed by flow cytometry. See Materials and Methods for full procedure. ( b–d ) Intracellular cytokine staining of OT-I cells in ( a ). Cells were treated with 10 μg/ml brefeldin A, incubated for 5 hours at 37 °C, stained with antibodies against CD25, interleukin (IL)-2, and interferon (IFN)-γ, and analyzed by flow cytometry. OT-I responses following incubation with min.2:OVA versus cntrl.36:OVA and NLDC145:OVA versus IgG2a:OVA were compared by student's t -test (**** P

    Techniques Used: Functional Assay, In Vitro, Labeling, Incubation, Isolation, Mouse Assay, Flow Cytometry, Cytometry, Staining

    13) Product Images from "Lipid raft-associated β-adducin participates in neutrophil migration"

    Article Title: Lipid raft-associated β-adducin participates in neutrophil migration

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2018.9113

    The role of the lipid raft structure in neutrophil migration. (A) Neutrophils were pre-adhered onto the coverslip coated with fibrinogen, and Iscove's modified Dulbecco's medium co-ntaining fMLP was added to one side of the coverslip. Confocal microscopy images showed the distribution of GM1 (lipid raft maker). GM1 was detected using AlexaFluor-488-conjugated CTxB. (B) Neutrophils were treated with 5 mM MβCD, then cells were treated as described above. Confocal microscopy images showed the distribution of GM1 (lipid raft maker). (C) MβCD (5 mM) was used to treat the peripheral blood neutrophils, in a Transwell migration assay. (D) Transwell assay for dHL-60 cells. Cell migratory rate was determined. Migratory rate=the number of migratory cells/the number of total cells. GM1, monosialotetrahexosylganglioside; fMLP, N-formylmethionyl-leucyl-phenyl-alanine; MβCD, methyl-β-cyclodextrin.
    Figure Legend Snippet: The role of the lipid raft structure in neutrophil migration. (A) Neutrophils were pre-adhered onto the coverslip coated with fibrinogen, and Iscove's modified Dulbecco's medium co-ntaining fMLP was added to one side of the coverslip. Confocal microscopy images showed the distribution of GM1 (lipid raft maker). GM1 was detected using AlexaFluor-488-conjugated CTxB. (B) Neutrophils were treated with 5 mM MβCD, then cells were treated as described above. Confocal microscopy images showed the distribution of GM1 (lipid raft maker). (C) MβCD (5 mM) was used to treat the peripheral blood neutrophils, in a Transwell migration assay. (D) Transwell assay for dHL-60 cells. Cell migratory rate was determined. Migratory rate=the number of migratory cells/the number of total cells. GM1, monosialotetrahexosylganglioside; fMLP, N-formylmethionyl-leucyl-phenyl-alanine; MβCD, methyl-β-cyclodextrin.

    Techniques Used: Migration, Modification, Confocal Microscopy, Transwell Migration Assay, Transwell Assay

    14) Product Images from "Derivation of Transgene‐Free Rat Induced Pluripotent Stem Cells Approximating the Quality of Embryonic Stem Cells"

    Article Title: Derivation of Transgene‐Free Rat Induced Pluripotent Stem Cells Approximating the Quality of Embryonic Stem Cells

    Journal: Stem Cells Translational Medicine

    doi: 10.5966/sctm.2015-0390

    Establishment of riPS cells and ES cells. (A): Derivation of rat ES cells from blastocyst outgrowth. We flushed E4.5 blastocysts from the uterus of pregnant Sprague Dawley rats (left). Scale bar = 100 μm. The blastocysts were plated on feeders in the 3i/Lif medium until the primary outgrowth attached at day 4. ES cells were established from blastocyst outgrowth after three passages and showed typical ES cell morphology. Scale bar = 200 μm. (B): Schematic diagram of the EBNA/oriP‐based reprogramming vector. Plasmid pMaster3 contains 7 human transcription factor genes of OCT4/POU5f1 , SOX2 , KLF4 , C‐MYC , NANOG , LIN28 , and NR5A2 for reprogramming and two drug resistance genes, neo and HSVtk , for positive/negative selection. Plasmid pMaster12 and pMaster22 with additional human and rat miR‐302/367 gene cluster, respectively. (C): Flowchart of transgene‐free riPS cell derivation. Rat fibroblast cells were seeded on mouse embryonic fibroblast feeders and cultured in serum medium after electrotransfection with pMaster3/pMaster12/pMaster22, followed by G418 drug selection for 5 days to ensure successful plasmid transfection. After the eighth day, 3i/Lif medium was used for riPS cell maintenance and primary clones were picked and propagated. To obtain transgene‐free subclones, we plated 20,000 cells into a 10‐cm dish to carry out FIAU negative selection to exclude cells carrying transgenes after the second day. Clones that passed negative selection were picked and expanded for further verification. G418, geneticin, a derivative of gentamycin; serum medium, Dulbecco's modified Eagle's medium supplemented with fetal bovine serum and Lif; and 3i/Lif medium, N2B27 based serum‐free medium with additional CHIR99021, PD0325901, A83‐01, and Lif. (D): Cell morphology during different reprogramming stages. Rat embryonic fibroblasts at passage 1 were used for reprogramming. Primary riPS cells have rat ES‐cell like morphology at day 10 after electrotransfection under hypoxic culture conditions in 3i/Lif medium. The established iPS clones showed round and smooth border morphology, which is a typical characteristic of ES cells. Scale bar = 200 μm. Abbreviations: ES, embryonic stem; FIAU, 1‐(2‐deoxy‐2‐fluoro‐1‐D‐arabinofuranosyl)‐5‐iodoracil; iPS, induced pluripotent stem; Lif, leukemia inhibitory factor; riPS, rat induced pluripotent stem.
    Figure Legend Snippet: Establishment of riPS cells and ES cells. (A): Derivation of rat ES cells from blastocyst outgrowth. We flushed E4.5 blastocysts from the uterus of pregnant Sprague Dawley rats (left). Scale bar = 100 μm. The blastocysts were plated on feeders in the 3i/Lif medium until the primary outgrowth attached at day 4. ES cells were established from blastocyst outgrowth after three passages and showed typical ES cell morphology. Scale bar = 200 μm. (B): Schematic diagram of the EBNA/oriP‐based reprogramming vector. Plasmid pMaster3 contains 7 human transcription factor genes of OCT4/POU5f1 , SOX2 , KLF4 , C‐MYC , NANOG , LIN28 , and NR5A2 for reprogramming and two drug resistance genes, neo and HSVtk , for positive/negative selection. Plasmid pMaster12 and pMaster22 with additional human and rat miR‐302/367 gene cluster, respectively. (C): Flowchart of transgene‐free riPS cell derivation. Rat fibroblast cells were seeded on mouse embryonic fibroblast feeders and cultured in serum medium after electrotransfection with pMaster3/pMaster12/pMaster22, followed by G418 drug selection for 5 days to ensure successful plasmid transfection. After the eighth day, 3i/Lif medium was used for riPS cell maintenance and primary clones were picked and propagated. To obtain transgene‐free subclones, we plated 20,000 cells into a 10‐cm dish to carry out FIAU negative selection to exclude cells carrying transgenes after the second day. Clones that passed negative selection were picked and expanded for further verification. G418, geneticin, a derivative of gentamycin; serum medium, Dulbecco's modified Eagle's medium supplemented with fetal bovine serum and Lif; and 3i/Lif medium, N2B27 based serum‐free medium with additional CHIR99021, PD0325901, A83‐01, and Lif. (D): Cell morphology during different reprogramming stages. Rat embryonic fibroblasts at passage 1 were used for reprogramming. Primary riPS cells have rat ES‐cell like morphology at day 10 after electrotransfection under hypoxic culture conditions in 3i/Lif medium. The established iPS clones showed round and smooth border morphology, which is a typical characteristic of ES cells. Scale bar = 200 μm. Abbreviations: ES, embryonic stem; FIAU, 1‐(2‐deoxy‐2‐fluoro‐1‐D‐arabinofuranosyl)‐5‐iodoracil; iPS, induced pluripotent stem; Lif, leukemia inhibitory factor; riPS, rat induced pluripotent stem.

    Techniques Used: Plasmid Preparation, Selection, Cell Culture, Transfection, Clone Assay, Modification

    15) Product Images from "Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration"

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration

    Journal: Toxins

    doi: 10.3390/toxins9040125

    Chemokine ELISArray profiles of human skin fibroblasts HFF-1 cells. Fibroblasts HFF-1 (1 × 10 5 cells/mL) cultures were treated with 5 μg/mL recombinant PLD of L. laeta (rLlPLD1, active) or (rLlPLD2, inactive), 10 μM ceramide 1-phosphate (C1P), 10 μM lysophosphatidic acid (LPA), or 10 μg/mL LPS and incubated for 24 h at 37 °C in an atmosphere containing 5% CO 2 . Dulbecco’s Modified Eagle’s Medium (DMEM) without fetal bovine serum (FBS) was used as control. Culture supernatants were recovered and used for chemokine screening in a Common Chemokines Multi-Analyte ELISArray kit (QIAGEN) panel for the detection of interleukin (IL)-8, monocyte chemoattractant protein-1 (MCP-1), RANTES, MIP-1α, MIP-1β, IP-10, I-TAC, MIG, Eotaxin, TARC, MDC, and GRO-α, according to the manufacturer’s instructions. Absorbance at 450 nm was measured in a micro-plate reader, and the chemokine levels were determined in relation to the absorbance value of the negative control (assay buffer) and compared to the positive control (containing standard of all 12 chemokines). The presence of chemokines in culture supernatants was considered for absorbance values over negative control. Results were expressed as mean ± SEM of two experiments performed independently.
    Figure Legend Snippet: Chemokine ELISArray profiles of human skin fibroblasts HFF-1 cells. Fibroblasts HFF-1 (1 × 10 5 cells/mL) cultures were treated with 5 μg/mL recombinant PLD of L. laeta (rLlPLD1, active) or (rLlPLD2, inactive), 10 μM ceramide 1-phosphate (C1P), 10 μM lysophosphatidic acid (LPA), or 10 μg/mL LPS and incubated for 24 h at 37 °C in an atmosphere containing 5% CO 2 . Dulbecco’s Modified Eagle’s Medium (DMEM) without fetal bovine serum (FBS) was used as control. Culture supernatants were recovered and used for chemokine screening in a Common Chemokines Multi-Analyte ELISArray kit (QIAGEN) panel for the detection of interleukin (IL)-8, monocyte chemoattractant protein-1 (MCP-1), RANTES, MIP-1α, MIP-1β, IP-10, I-TAC, MIG, Eotaxin, TARC, MDC, and GRO-α, according to the manufacturer’s instructions. Absorbance at 450 nm was measured in a micro-plate reader, and the chemokine levels were determined in relation to the absorbance value of the negative control (assay buffer) and compared to the positive control (containing standard of all 12 chemokines). The presence of chemokines in culture supernatants was considered for absorbance values over negative control. Results were expressed as mean ± SEM of two experiments performed independently.

    Techniques Used: Recombinant, Incubation, Modification, Negative Control, Positive Control

    16) Product Images from "Modulation of adipose tissue lipolysis and body weight by high-density lipoproteins in mice"

    Article Title: Modulation of adipose tissue lipolysis and body weight by high-density lipoproteins in mice

    Journal: Nutrition & Diabetes

    doi: 10.1038/nutd.2014.4

    HDL promotes catecholamine but not cAMP-stimulated lipolysis in 3T3-L1 adipocytes. 3T3-L1 adipocytes at day 10 after differentiation were cultured in Dulbecco's modified Eagle's medium containing fatty acid-free 0.2% bovine serum albumin for 24 h, and followed by incubation with or without lipolytic stimuli, 5 μ M ISOP or 500 m M 8-Br-cAMP, and with or without human HDL and apoA-I in indicated concentrations for 4 h. Net release of glycerol into conditioned media was measured as the indicator of lipolysis. Whole-cell lysates were prepared and immunoblotted for p-HSL, HSL and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as described in Materials and methods section. ( a ) Basal lipolysis of 3T3-L1 adipocytes with HDL and apoA-I. ( b ) Catecholamine ISOP-stimulated lipolysis with HDL and apoA1. ( c ) cAMP-stimulated lipolysis with HDL and apoA-I. ( d ) Immunoblots of p-HSL, HSL and GAPDH in 3T3-L1 adipocytes under ISOP stimulation with HDL and apoA-I (upper panel). Densitometric data for p-HSL/HSL (lower panel). The image is one representative immunoblot from three independent experiments. The asterisk denotes statistically significant differences compared with control (analysis of variance (ANOVA), P
    Figure Legend Snippet: HDL promotes catecholamine but not cAMP-stimulated lipolysis in 3T3-L1 adipocytes. 3T3-L1 adipocytes at day 10 after differentiation were cultured in Dulbecco's modified Eagle's medium containing fatty acid-free 0.2% bovine serum albumin for 24 h, and followed by incubation with or without lipolytic stimuli, 5 μ M ISOP or 500 m M 8-Br-cAMP, and with or without human HDL and apoA-I in indicated concentrations for 4 h. Net release of glycerol into conditioned media was measured as the indicator of lipolysis. Whole-cell lysates were prepared and immunoblotted for p-HSL, HSL and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as described in Materials and methods section. ( a ) Basal lipolysis of 3T3-L1 adipocytes with HDL and apoA-I. ( b ) Catecholamine ISOP-stimulated lipolysis with HDL and apoA1. ( c ) cAMP-stimulated lipolysis with HDL and apoA-I. ( d ) Immunoblots of p-HSL, HSL and GAPDH in 3T3-L1 adipocytes under ISOP stimulation with HDL and apoA-I (upper panel). Densitometric data for p-HSL/HSL (lower panel). The image is one representative immunoblot from three independent experiments. The asterisk denotes statistically significant differences compared with control (analysis of variance (ANOVA), P

    Techniques Used: Cell Culture, Modification, Incubation, Western Blot

    17) Product Images from "Extracellular Signals of a Human Epithelial Colorectal Adenocarcinoma (Caco-2) Cell Line Facilitate the Penetration of Pseudomonas aeruginosa PAO1 Strain through the Mucin Layer"

    Article Title: Extracellular Signals of a Human Epithelial Colorectal Adenocarcinoma (Caco-2) Cell Line Facilitate the Penetration of Pseudomonas aeruginosa PAO1 Strain through the Mucin Layer

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2017.00415

    GRO-α facilitates a flagellar filament rotation of the P. aeruginosa PAO1 strain. P. aeruginosa PAO1 strain cells were loaded onto a glass slide precoated with flagellar filament FliC antibodies, and growth regulated oncogene-α or Dulbecco's modified Eagle medium (DMEM; control) was added. Bacterial cells were visualized and recorded as a movie using EVOS microscope, showing videos of the tethered bacteria. The rotational speed was calculated from the number of rotations in 60 s. Error bars indicate standard error ( n = 10). * P
    Figure Legend Snippet: GRO-α facilitates a flagellar filament rotation of the P. aeruginosa PAO1 strain. P. aeruginosa PAO1 strain cells were loaded onto a glass slide precoated with flagellar filament FliC antibodies, and growth regulated oncogene-α or Dulbecco's modified Eagle medium (DMEM; control) was added. Bacterial cells were visualized and recorded as a movie using EVOS microscope, showing videos of the tethered bacteria. The rotational speed was calculated from the number of rotations in 60 s. Error bars indicate standard error ( n = 10). * P

    Techniques Used: Modification, Microscopy

    GRO-α facilitates a swarming motility of the P. aeruginosa PAO1 strain. (A) The P. aeruginosa PAO1 strain was spotted on swarming agar containing growth regulated oncogene -α or Dulbecco's modified Eagle medium (DMEM; control). After incubation for 14 h, the swarming agar plate was observed and photographed. (B) The radial distance (mm) from the center of the agar was measured. Error bars indicate standard error ( n = 3). * P
    Figure Legend Snippet: GRO-α facilitates a swarming motility of the P. aeruginosa PAO1 strain. (A) The P. aeruginosa PAO1 strain was spotted on swarming agar containing growth regulated oncogene -α or Dulbecco's modified Eagle medium (DMEM; control). After incubation for 14 h, the swarming agar plate was observed and photographed. (B) The radial distance (mm) from the center of the agar was measured. Error bars indicate standard error ( n = 3). * P

    Techniques Used: Modification, Incubation

    GRO-α does not attract the P. aeruginosa PAO1 strain. (A) A glass slide was filled with the GFP-expressing P. aeruginosa PAO1 strain. A capillary tube was filled with growth regulated oncogene-α or Dulbecco's modified Eagle medium (DMEM; control). The accumulation of P. aeruginosa at the capillary tip was visualized and photographed by microscopy at a magnification of 40×. (B) The graph shows the relative intensity of GFP in the capillary tube after incubation for 10 min. Error bars indicate standard error ( n = 3).
    Figure Legend Snippet: GRO-α does not attract the P. aeruginosa PAO1 strain. (A) A glass slide was filled with the GFP-expressing P. aeruginosa PAO1 strain. A capillary tube was filled with growth regulated oncogene-α or Dulbecco's modified Eagle medium (DMEM; control). The accumulation of P. aeruginosa at the capillary tip was visualized and photographed by microscopy at a magnification of 40×. (B) The graph shows the relative intensity of GFP in the capillary tube after incubation for 10 min. Error bars indicate standard error ( n = 3).

    Techniques Used: Expressing, Modification, Microscopy, Incubation

    Characterization of Caco-2 cell supernatant facilitating penetration of P. aeruginosa PAO1 strain through the mucin layer. A 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control) were filtered through 10- or 3-kDa MW cut-off membranes and treated with or without a trypsin (trypsin + or −). The filtered and unfiltered samples with or without trypsin filled the bottom chambers of Transwell. The P. aeruginosa PAO1 strain was added to the top chambers, and after 3 h, the numbers of bacteria in the bottom chambers were counted. Error bars indicate standard error ( n = 3). * P
    Figure Legend Snippet: Characterization of Caco-2 cell supernatant facilitating penetration of P. aeruginosa PAO1 strain through the mucin layer. A 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control) were filtered through 10- or 3-kDa MW cut-off membranes and treated with or without a trypsin (trypsin + or −). The filtered and unfiltered samples with or without trypsin filled the bottom chambers of Transwell. The P. aeruginosa PAO1 strain was added to the top chambers, and after 3 h, the numbers of bacteria in the bottom chambers were counted. Error bars indicate standard error ( n = 3). * P

    Techniques Used: Modification

    Caco-2 cell supernatant facilitates a swarming motility of the P. aeruginosa PAO1 strain. (A) The P. aeruginosa PAO1 strain was incubated in Caco-2 cell supernatant (Caco-2 sup.) or Dulbecco's modified Eagle medium (DMEM; control) at 37°C in 5% CO 2 . After incubation, the optical density (600 nm) of the culture was measured at the indicated times. Error bars indicate standard error ( n = 3). (B) After incubation of the P. aeruginosa PAO1 strain in Caco-2 cell supernatant (Caco-2 sup.) or DMEM (control) at 37°C in 5% CO 2 for 5 h, the azocasein degradation activity of the collected culture supernatant was measured. Error bars indicate standard error ( n = 3). (C) A 15-day Caco-2 cell supernatant (Caco-2 sup.) and DMEM (control) were filtered through 10-kDa MW cut-off membranes. The
    Figure Legend Snippet: Caco-2 cell supernatant facilitates a swarming motility of the P. aeruginosa PAO1 strain. (A) The P. aeruginosa PAO1 strain was incubated in Caco-2 cell supernatant (Caco-2 sup.) or Dulbecco's modified Eagle medium (DMEM; control) at 37°C in 5% CO 2 . After incubation, the optical density (600 nm) of the culture was measured at the indicated times. Error bars indicate standard error ( n = 3). (B) After incubation of the P. aeruginosa PAO1 strain in Caco-2 cell supernatant (Caco-2 sup.) or DMEM (control) at 37°C in 5% CO 2 for 5 h, the azocasein degradation activity of the collected culture supernatant was measured. Error bars indicate standard error ( n = 3). (C) A 15-day Caco-2 cell supernatant (Caco-2 sup.) and DMEM (control) were filtered through 10-kDa MW cut-off membranes. The

    Techniques Used: Incubation, Modification, Activity Assay

    Caco-2 cell supernatant facilitates a flagellar filament rotation of the P. aeruginosa PAO1 strain. A 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control) were filtered through 10-kDa MW cut-off membranes. The
    Figure Legend Snippet: Caco-2 cell supernatant facilitates a flagellar filament rotation of the P. aeruginosa PAO1 strain. A 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control) were filtered through 10-kDa MW cut-off membranes. The

    Techniques Used: Modification

    Chemokines secreted by Caco-2 cells facilitate the penetration of the P. aeruginosa PAO1 strain through the mucin layer. (A) Cytokine levels in the 15-day Caco-2 cell supernatant were determined by enzyme-linked immunosorbent assay (ELISA). The graph shows the cytokine concentrations in the supernatant. Error bars indicate standard error ( n = 3). (B) The cytokines identified by the assay were placed in the bottom chamber of Transwell, as were unfiltered 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control). The P. aeruginosa PAO1 strain was added to the top chambers and, after 3 h, the numbers of bacteria in the bottom chamber were counted. Error bars indicate standard error ( n = 3). * P
    Figure Legend Snippet: Chemokines secreted by Caco-2 cells facilitate the penetration of the P. aeruginosa PAO1 strain through the mucin layer. (A) Cytokine levels in the 15-day Caco-2 cell supernatant were determined by enzyme-linked immunosorbent assay (ELISA). The graph shows the cytokine concentrations in the supernatant. Error bars indicate standard error ( n = 3). (B) The cytokines identified by the assay were placed in the bottom chamber of Transwell, as were unfiltered 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control). The P. aeruginosa PAO1 strain was added to the top chambers and, after 3 h, the numbers of bacteria in the bottom chamber were counted. Error bars indicate standard error ( n = 3). * P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Modification

    Caco-2 cell supernatant facilitates penetration of the P. aeruginosa PAO1 strain through the mucin layer. (A) The bottom chambers of Transwell cells were filled with Caco-2 cell supernatant (collected after 5, 10, and 15 days of culture) or with Dulbecco's modified Eagle medium (DMEM; control). The P. aeruginosa PAO1 strain was added to the top chamber. After 3 h, the numbers of bacteria in the bottom chambers were counted. Error bars indicate standard error ( n = 3). * P
    Figure Legend Snippet: Caco-2 cell supernatant facilitates penetration of the P. aeruginosa PAO1 strain through the mucin layer. (A) The bottom chambers of Transwell cells were filled with Caco-2 cell supernatant (collected after 5, 10, and 15 days of culture) or with Dulbecco's modified Eagle medium (DMEM; control). The P. aeruginosa PAO1 strain was added to the top chamber. After 3 h, the numbers of bacteria in the bottom chambers were counted. Error bars indicate standard error ( n = 3). * P

    Techniques Used: Modification

    18) Product Images from "Pharmacological disruption of insulin-like growth factor 1 binding to IGF-binding proteins restores anabolic responses in human osteoarthritic chondrocytes"

    Article Title: Pharmacological disruption of insulin-like growth factor 1 binding to IGF-binding proteins restores anabolic responses in human osteoarthritic chondrocytes

    Journal: Arthritis Research & Therapy

    doi: 10.1186/ar1201

    Competition of NBI-31772, a small-molecule inhibitor of the binding of insulin-like growth factor (IGF)-1 to IGF-binding proteins (IGFBPs), with 125 I-labeled IGF-1 binding to cartilage. Explants cut from a homogeneous area of osteoarthritic cartilage were separated in equal groups and incubated with 11.7 nM R 3 IGF-1 (an analog of IGF-1 with a greatly reduced affinity for IGFBPs), 11.7 nM IGF-1, 1.5 M NaCl or 10 μM NBI-31772 in 1 ml of Dulbecco's phosphate-buffered saline (DPBS) for 24 hours at 4°C. 125 I-labeled IGF-1 (20,000 dpm) was added to each incubation medium and the cells were incubated for 24 more hours. The medium was harvested and explants were washed three times with DPBS. The radioactivity in medium, lavage buffer, and cartilage explants was counted. The percentage of 125 I-labeled IGF-1 remaining in cartilage was corrected for the weight of explants and divided by total counts. Data are the means of two experiments. Results are expressed as percentages of the control value (cartilage incubated with 125 I-labeled IGF-1 alone).
    Figure Legend Snippet: Competition of NBI-31772, a small-molecule inhibitor of the binding of insulin-like growth factor (IGF)-1 to IGF-binding proteins (IGFBPs), with 125 I-labeled IGF-1 binding to cartilage. Explants cut from a homogeneous area of osteoarthritic cartilage were separated in equal groups and incubated with 11.7 nM R 3 IGF-1 (an analog of IGF-1 with a greatly reduced affinity for IGFBPs), 11.7 nM IGF-1, 1.5 M NaCl or 10 μM NBI-31772 in 1 ml of Dulbecco's phosphate-buffered saline (DPBS) for 24 hours at 4°C. 125 I-labeled IGF-1 (20,000 dpm) was added to each incubation medium and the cells were incubated for 24 more hours. The medium was harvested and explants were washed three times with DPBS. The radioactivity in medium, lavage buffer, and cartilage explants was counted. The percentage of 125 I-labeled IGF-1 remaining in cartilage was corrected for the weight of explants and divided by total counts. Data are the means of two experiments. Results are expressed as percentages of the control value (cartilage incubated with 125 I-labeled IGF-1 alone).

    Techniques Used: Binding Assay, Labeling, Incubation, Radioactivity

    Effect of NBI-31772, a small-molecule inhibitor of the binding of insulin-like growth factor (IGF)-1 to IGF-binding proteins, on 125 I-labeled IGF-1 binding to human osteoarthritic chondrocytes. Confluent chondrocytes were incubated with 1 × 10 5 dpm 125 I-labeled IGF-1 with or without excess (150 nM) IGF-1 or 10 μM NBI-31772 for 1 hour at 4°C. Medium was harvested and cells were washed three times with Dulbecco's phosphate-buffered saline. Cells were lysed in 1 N NaOH. The percentage of 125 I-labeled IGF-1 bound to the cells was calculated as dpm in NaOH-treated cells over total dpm. Data are expressed as the means and SEM of four replicates from a representative experiment (out of two independent cultures).
    Figure Legend Snippet: Effect of NBI-31772, a small-molecule inhibitor of the binding of insulin-like growth factor (IGF)-1 to IGF-binding proteins, on 125 I-labeled IGF-1 binding to human osteoarthritic chondrocytes. Confluent chondrocytes were incubated with 1 × 10 5 dpm 125 I-labeled IGF-1 with or without excess (150 nM) IGF-1 or 10 μM NBI-31772 for 1 hour at 4°C. Medium was harvested and cells were washed three times with Dulbecco's phosphate-buffered saline. Cells were lysed in 1 N NaOH. The percentage of 125 I-labeled IGF-1 bound to the cells was calculated as dpm in NaOH-treated cells over total dpm. Data are expressed as the means and SEM of four replicates from a representative experiment (out of two independent cultures).

    Techniques Used: Binding Assay, Labeling, Incubation

    19) Product Images from "Serum-mediated Activation of Bone Marrow–derived Mesenchymal Stem Cells in Ischemic Stroke Patients"

    Article Title: Serum-mediated Activation of Bone Marrow–derived Mesenchymal Stem Cells in Ischemic Stroke Patients

    Journal: Cell Transplantation

    doi: 10.1177/0963689718755404

    Evaluation of phenotypic characteristics of mesenchymal stem cells (MSCs). (A) Representative phase contrast images of human MSCs (hMSCs) expanded with the different serums. (B) Cumulative population doubling level of hMSCs cultured in Dulbecco’s modified Eagle’s medium with 10% fetal bovine serum (FBS), control serum (NS), and stroke patient serum (SS). (C) Fluorescent-activated cell sorting analysis of hMSCs cultured with different types of serum. Quantitative analysis of the percentages of cells expressing CD90, CD73 (positive markers), and CD34, CD45 (negative markers). The relative expression levels of both human vascular endothelial growth factor (D) and human fibroblast growth factor (F) were significantly increased in SS-hMSCs than FBS-hMSCs or NS-hMSCs. Human glial cell–derived neurotrophic factor (E) expression level was significantly lower in FBS-hMSCs than NS-hMSCs and SS-hMSCs. All data are presented as mean + SD (** P < 0.01, n = 6).
    Figure Legend Snippet: Evaluation of phenotypic characteristics of mesenchymal stem cells (MSCs). (A) Representative phase contrast images of human MSCs (hMSCs) expanded with the different serums. (B) Cumulative population doubling level of hMSCs cultured in Dulbecco’s modified Eagle’s medium with 10% fetal bovine serum (FBS), control serum (NS), and stroke patient serum (SS). (C) Fluorescent-activated cell sorting analysis of hMSCs cultured with different types of serum. Quantitative analysis of the percentages of cells expressing CD90, CD73 (positive markers), and CD34, CD45 (negative markers). The relative expression levels of both human vascular endothelial growth factor (D) and human fibroblast growth factor (F) were significantly increased in SS-hMSCs than FBS-hMSCs or NS-hMSCs. Human glial cell–derived neurotrophic factor (E) expression level was significantly lower in FBS-hMSCs than NS-hMSCs and SS-hMSCs. All data are presented as mean + SD (** P < 0.01, n = 6).

    Techniques Used: Cell Culture, Modification, FACS, Expressing, Derivative Assay

    20) Product Images from "Extracellular Signals of a Human Epithelial Colorectal Adenocarcinoma (Caco-2) Cell Line Facilitate the Penetration of Pseudomonas aeruginosa PAO1 Strain through the Mucin Layer"

    Article Title: Extracellular Signals of a Human Epithelial Colorectal Adenocarcinoma (Caco-2) Cell Line Facilitate the Penetration of Pseudomonas aeruginosa PAO1 Strain through the Mucin Layer

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2017.00415

    GRO-α facilitates a flagellar filament rotation of the P. aeruginosa PAO1 strain. P. aeruginosa PAO1 strain cells were loaded onto a glass slide precoated with flagellar filament FliC antibodies, and growth regulated oncogene-α or Dulbecco's modified Eagle medium (DMEM; control) was added. Bacterial cells were visualized and recorded as a movie using EVOS microscope, showing videos of the tethered bacteria. The rotational speed was calculated from the number of rotations in 60 s. Error bars indicate standard error ( n = 10). * P
    Figure Legend Snippet: GRO-α facilitates a flagellar filament rotation of the P. aeruginosa PAO1 strain. P. aeruginosa PAO1 strain cells were loaded onto a glass slide precoated with flagellar filament FliC antibodies, and growth regulated oncogene-α or Dulbecco's modified Eagle medium (DMEM; control) was added. Bacterial cells were visualized and recorded as a movie using EVOS microscope, showing videos of the tethered bacteria. The rotational speed was calculated from the number of rotations in 60 s. Error bars indicate standard error ( n = 10). * P

    Techniques Used: Modification, Microscopy

    GRO-α facilitates a swarming motility of the P. aeruginosa PAO1 strain. (A) The P. aeruginosa PAO1 strain was spotted on swarming agar containing growth regulated oncogene -α or Dulbecco's modified Eagle medium (DMEM; control). After incubation for 14 h, the swarming agar plate was observed and photographed. (B) The radial distance (mm) from the center of the agar was measured. Error bars indicate standard error ( n = 3). * P
    Figure Legend Snippet: GRO-α facilitates a swarming motility of the P. aeruginosa PAO1 strain. (A) The P. aeruginosa PAO1 strain was spotted on swarming agar containing growth regulated oncogene -α or Dulbecco's modified Eagle medium (DMEM; control). After incubation for 14 h, the swarming agar plate was observed and photographed. (B) The radial distance (mm) from the center of the agar was measured. Error bars indicate standard error ( n = 3). * P

    Techniques Used: Modification, Incubation

    GRO-α does not attract the P. aeruginosa PAO1 strain. (A) A glass slide was filled with the GFP-expressing P. aeruginosa PAO1 strain. A capillary tube was filled with growth regulated oncogene-α or Dulbecco's modified Eagle medium (DMEM; control). The accumulation of P. aeruginosa at the capillary tip was visualized and photographed by microscopy at a magnification of 40×. (B) The graph shows the relative intensity of GFP in the capillary tube after incubation for 10 min. Error bars indicate standard error ( n = 3).
    Figure Legend Snippet: GRO-α does not attract the P. aeruginosa PAO1 strain. (A) A glass slide was filled with the GFP-expressing P. aeruginosa PAO1 strain. A capillary tube was filled with growth regulated oncogene-α or Dulbecco's modified Eagle medium (DMEM; control). The accumulation of P. aeruginosa at the capillary tip was visualized and photographed by microscopy at a magnification of 40×. (B) The graph shows the relative intensity of GFP in the capillary tube after incubation for 10 min. Error bars indicate standard error ( n = 3).

    Techniques Used: Expressing, Modification, Microscopy, Incubation

    Characterization of Caco-2 cell supernatant facilitating penetration of P. aeruginosa PAO1 strain through the mucin layer. A 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control) were filtered through 10- or 3-kDa MW cut-off membranes and treated with or without a trypsin (trypsin + or −). The filtered and unfiltered samples with or without trypsin filled the bottom chambers of Transwell. The P. aeruginosa PAO1 strain was added to the top chambers, and after 3 h, the numbers of bacteria in the bottom chambers were counted. Error bars indicate standard error ( n = 3). * P
    Figure Legend Snippet: Characterization of Caco-2 cell supernatant facilitating penetration of P. aeruginosa PAO1 strain through the mucin layer. A 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control) were filtered through 10- or 3-kDa MW cut-off membranes and treated with or without a trypsin (trypsin + or −). The filtered and unfiltered samples with or without trypsin filled the bottom chambers of Transwell. The P. aeruginosa PAO1 strain was added to the top chambers, and after 3 h, the numbers of bacteria in the bottom chambers were counted. Error bars indicate standard error ( n = 3). * P

    Techniques Used: Modification

    Caco-2 cell supernatant facilitates a swarming motility of the P. aeruginosa PAO1 strain. (A) The P. aeruginosa PAO1 strain was incubated in Caco-2 cell supernatant (Caco-2 sup.) or Dulbecco's modified Eagle medium (DMEM; control) at 37°C in 5% CO 2 . After incubation, the optical density (600 nm) of the culture was measured at the indicated times. Error bars indicate standard error ( n = 3). (B) After incubation of the P. aeruginosa PAO1 strain in Caco-2 cell supernatant (Caco-2 sup.) or DMEM (control) at 37°C in 5% CO 2 for 5 h, the azocasein degradation activity of the collected culture supernatant was measured. Error bars indicate standard error ( n = 3). (C) A 15-day Caco-2 cell supernatant (Caco-2 sup.) and DMEM (control) were filtered through 10-kDa MW cut-off membranes. The
    Figure Legend Snippet: Caco-2 cell supernatant facilitates a swarming motility of the P. aeruginosa PAO1 strain. (A) The P. aeruginosa PAO1 strain was incubated in Caco-2 cell supernatant (Caco-2 sup.) or Dulbecco's modified Eagle medium (DMEM; control) at 37°C in 5% CO 2 . After incubation, the optical density (600 nm) of the culture was measured at the indicated times. Error bars indicate standard error ( n = 3). (B) After incubation of the P. aeruginosa PAO1 strain in Caco-2 cell supernatant (Caco-2 sup.) or DMEM (control) at 37°C in 5% CO 2 for 5 h, the azocasein degradation activity of the collected culture supernatant was measured. Error bars indicate standard error ( n = 3). (C) A 15-day Caco-2 cell supernatant (Caco-2 sup.) and DMEM (control) were filtered through 10-kDa MW cut-off membranes. The

    Techniques Used: Incubation, Modification, Activity Assay

    Caco-2 cell supernatant facilitates a flagellar filament rotation of the P. aeruginosa PAO1 strain. A 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control) were filtered through 10-kDa MW cut-off membranes. The
    Figure Legend Snippet: Caco-2 cell supernatant facilitates a flagellar filament rotation of the P. aeruginosa PAO1 strain. A 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control) were filtered through 10-kDa MW cut-off membranes. The

    Techniques Used: Modification

    Chemokines secreted by Caco-2 cells facilitate the penetration of the P. aeruginosa PAO1 strain through the mucin layer. (A) Cytokine levels in the 15-day Caco-2 cell supernatant were determined by enzyme-linked immunosorbent assay (ELISA). The graph shows the cytokine concentrations in the supernatant. Error bars indicate standard error ( n = 3). (B) The cytokines identified by the assay were placed in the bottom chamber of Transwell, as were unfiltered 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control). The P. aeruginosa PAO1 strain was added to the top chambers and, after 3 h, the numbers of bacteria in the bottom chamber were counted. Error bars indicate standard error ( n = 3). * P
    Figure Legend Snippet: Chemokines secreted by Caco-2 cells facilitate the penetration of the P. aeruginosa PAO1 strain through the mucin layer. (A) Cytokine levels in the 15-day Caco-2 cell supernatant were determined by enzyme-linked immunosorbent assay (ELISA). The graph shows the cytokine concentrations in the supernatant. Error bars indicate standard error ( n = 3). (B) The cytokines identified by the assay were placed in the bottom chamber of Transwell, as were unfiltered 15-day Caco-2 cell supernatant and Dulbecco's modified Eagle medium (DMEM; control). The P. aeruginosa PAO1 strain was added to the top chambers and, after 3 h, the numbers of bacteria in the bottom chamber were counted. Error bars indicate standard error ( n = 3). * P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Modification

    Caco-2 cell supernatant facilitates penetration of the P. aeruginosa PAO1 strain through the mucin layer. (A) The bottom chambers of Transwell cells were filled with Caco-2 cell supernatant (collected after 5, 10, and 15 days of culture) or with Dulbecco's modified Eagle medium (DMEM; control). The P. aeruginosa PAO1 strain was added to the top chamber. After 3 h, the numbers of bacteria in the bottom chambers were counted. Error bars indicate standard error ( n = 3). * P
    Figure Legend Snippet: Caco-2 cell supernatant facilitates penetration of the P. aeruginosa PAO1 strain through the mucin layer. (A) The bottom chambers of Transwell cells were filled with Caco-2 cell supernatant (collected after 5, 10, and 15 days of culture) or with Dulbecco's modified Eagle medium (DMEM; control). The P. aeruginosa PAO1 strain was added to the top chamber. After 3 h, the numbers of bacteria in the bottom chambers were counted. Error bars indicate standard error ( n = 3). * P

    Techniques Used: Modification

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    Article Snippet: The cells were grown in culture media containing 1× Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Rockville, MD, USA), conditioned medium, supplemented with 10% fetal calf serum (FCS; HyClone, Logan, UT, USA), and 5 μg/mL plasmocin™ (InvivoGen, San Diego, CA, USA) in a 5% CO2 incubator at 37°C. .. MCF-7 and MDA-MB-231 cell lines resistant against 5 μM and 10 μM tamoxifen were established in culture to gradual increases in concentrations of the indicated drugs in 1× DMEM conditioned medium.

    Article Title: Copper as an alternative antimicrobial coating for implants - An in vitro study
    Article Snippet: The concentration of copper released from the samples was measured by atomic absorption spectrometry (AAS) (ZEEnit 650, Analytik Jena AG, Jena, Germany) with electro-thermal atomization as described earlier[ ]. .. Briefly, the substrates were stored in 1 mL Dulbecco’s modified Eagle medium (DMEM, Invitrogen, Darmstadt, Germany) with 10% fetal calf serum (FCS, Superior, Biochrome, Berlin, Germany) and 1% gentamicin (Ratiopharm, Ulm, Germany) at 37 °C in a humidified atmosphere with 5% CO2 .

    Incubation:

    Article Title: Mesenchymal stem cell-derived extracellular vesicles attenuate influenza virus-induced acute lung injury in a pig model
    Article Snippet: Briefly, the tip of each bone was removed and the marrow was harvested by inserting a syringe needle into one end of the bone and flushing with Dulbecco’s modified Eagle’s medium (DMEM; Gibco). .. Cells (1–5 × 105 /cm2 ) were plated in 75-cm2 cell culture flasks in DMEM containing 10% fetal bovine serum (FBS; Gibco) and 1% antibiotic solution (Gibco) (C-DMEM).

    Article Title: Effect of fibroblast growth factor 9 on the osteogenic differentiation of bone marrow stromal stem cells and dental pulp stem cells
    Article Snippet: Following cleaning of the teeth, the pulp chambers were accessed by cutting the cementoenamel junction with a sterile fissure dental bur. .. Following exposure of the pulp, the pulp tissue was removed in fragments of ~0.5 mm, which were then placed onto a 6-cm culture dish containing Dulbecco’s modified Eagle’s medium (DMEM; Gibco-BRL, Grand Island, NY, USA) supplemented with 20% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) and antibiotics (Gibco-BRL) prior to incubation at 37°C in 5% CO2 . .. At confluence, the outgrown cells were transferred onto a 10-cm dish and then continuously passaged for further experiments.

    Article Title: Pharmacological disruption of insulin-like growth factor 1 binding to IGF-binding proteins restores anabolic responses in human osteoarthritic chondrocytes
    Article Snippet: Confluent cells were incubated in serum-free medium for 24 h. This medium was discarded and cells were further incubated in fresh, serum-free medium. .. Explants were dissected out from cartilage, washed extensively with Dulbecco's phosphate-buffered saline without calcium and magnesium (DPBS, Gibco), and incubated directly in serum-free Ham F12 medium without prior incubation in FCS-containing medium. .. Secretion media from chondrocytes or cartilage explants were collected after 48 hours and supplemented with protease inhibitors.

    Article Title: Effects of bradykinin on TGF-β1-induced epithelial-mesenchymal transition in ARPE-19 cells
    Article Snippet: ARPE-19 cells were cultured in Dulbecco's modified Eagle's medium (DMEM)/H (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.). .. ARPE-19 cells were cultured in Dulbecco's modified Eagle's medium (DMEM)/H (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.).

    Transfection:

    Article Title: Transforming growth factor-β decreases side population cells in hepatocellular carcinoma in vitro
    Article Snippet: Huh-7 and Huh-Bat [Huh-7 transfected with human Na+ -taurocholate co-transporting polypeptide (bile acid transporter) cDNA] cell lines were obtained from the Korean Cell Line Bank (Seoul, Korea). .. Cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Inc.).

    Activity Assay:

    Article Title: Laminarin-induced apoptosis in human colon cancer LoVo cells
    Article Snippet: In addition, this study may increase the development and application of laminarin for colon cancer treatment. .. The following reagents were used: Laminarin and paraformaldehyde (Sigma-Aldrich, St. Louis, MO, USA); hydroxycamptothecin (HCPT, Harbin Shengtai Pharmaceutical Co., Ltd., Harbin, China); Dulbecco’s modified Eagle’s medium (DMEM)/F12 culture medium (Thermo Fisher Scientific Inc., Waltham, MA, USA); fetal bovine serum (FBS; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China); pancreatin (Gibco-BRL, Rockville, MD, USA); Hoechst 33258 (Sigma-Aldrich); rabbit anti-human β-actin polyclonal antibody, DR4, DR5, TNF-related apoptosis-inducing ligand (TRAIL), Fas-associated protein with death domain (FADD) and Bid (Biosynthesis Biotechnology Co., Ltd., Beijing, China); mouse anti-human Bcl-2, rabbit anti-human Bax and fluorescein isothiocyanate (FITC)-goat anti-mouse polyclonal antibodies (Boster Biological Technology Co., Ltd., Wuhan, China); mouse anti-human caspase-8 and -3, alkaline phosphatase goat anti-rabbit polyclonal antibodies, as well as caspase-8, -3, -6 and -7 activity assay kits, sodium dodecyl sulphate sample buffer, polyacrylamide gel, nitrocellulose membranes, 5-bromo-4-chloro-3-indolyl-phosphate (BCIP)/nitroblue tetrazolium (NBT) alkaline phosphatase color development kit, Triton X-100, bovine serum albumin and phosphate-buffered saline (PBS; Beyotime Institute of Biotechnology, Haimen, China); detergent-compatible protein assay kit (Bio-Rad, Hercules, CA, USA). .. The CKX41 fluorescence inverted microscope was purchased from Olympus (Tokyo, Japan), and the Mini-Protean Tetra and Mini Trans-Blot electrophoresis systems, Gel Doc XR imaging system and Model 680 microplate reader were purchased from Bio-Rad.

    Article Title: MMP‐2 and MMP‐14 Silencing Inhibits VEGFR2 Cleavage and Induces the Differentiation of Porcine Adipose‐Derived Mesenchymal Stem Cells to Endothelial Cells
    Article Snippet: The adipose tissue from the abdominal fat was collected from the slaughterhouse and transferred to the laboratory in sterile conditions in Dulbecco's modified Eagle medium (DMEM; Invitrogen, Grand Island, NY, http://www.invitrogen.com ) with antibiotics 100 mg/ml penicillin (Sigma‐Aldrich, St. Louis, MO, http://www.sigmaaldrich.com ), and 100 mg/ml streptomycin (Sigma‐Aldrich). .. The adipose tissue from the abdominal fat was collected from the slaughterhouse and transferred to the laboratory in sterile conditions in Dulbecco's modified Eagle medium (DMEM; Invitrogen, Grand Island, NY, http://www.invitrogen.com ) with antibiotics 100 mg/ml penicillin (Sigma‐Aldrich, St. Louis, MO, http://www.sigmaaldrich.com ), and 100 mg/ml streptomycin (Sigma‐Aldrich).

    Microscopy:

    Article Title: Extracellular Signals of a Human Epithelial Colorectal Adenocarcinoma (Caco-2) Cell Line Facilitate the Penetration of Pseudomonas aeruginosa PAO1 Strain through the Mucin Layer
    Article Snippet: A glass slide was coated with Protein A (Biovision, Mountain View, CA, USA) for 30 min and washed with Dulbecco's phosphate buffered saline (DPBS; Gibco) to remove non-adherent Protein A. .. A glass slide was coated with Protein A (Biovision, Mountain View, CA, USA) for 30 min and washed with Dulbecco's phosphate buffered saline (DPBS; Gibco) to remove non-adherent Protein A.

    Modification:

    Article Title: Extracellular Signals of a Human Epithelial Colorectal Adenocarcinoma (Caco-2) Cell Line Facilitate the Penetration of Pseudomonas aeruginosa PAO1 Strain through the Mucin Layer
    Article Snippet: The Caco-2 cells were routinely grown at 37°C in a 95% air–5% CO2 atmosphere in Dulbecco's modified Eagle medium (DMEM) with high glucose (Sigma-Aldrich Co) and 10% heat-inactivated (56°C, 30 min) fetal bovine serum (FBS; Gibco, Grand Island, NY, USA). .. The cells were collected from the cell suspension by a centrifugation, and were washed with Dulbecco's phosphate buffered saline (DPBS; Gibco).

    Article Title: Mesenchymal stem cell-derived extracellular vesicles attenuate influenza virus-induced acute lung injury in a pig model
    Article Snippet: MSCs from femur bones of 2- to 6-week-old commercial pigs were isolated as described previously [ , ]. .. Briefly, the tip of each bone was removed and the marrow was harvested by inserting a syringe needle into one end of the bone and flushing with Dulbecco’s modified Eagle’s medium (DMEM; Gibco). .. The BM cells were filtered through a 70-μm nylon mesh filter (BD, Falcon, USA) and mononuclear cells were obtained by density gradient centrifugation over Ficoll-Hypaque.

    Article Title: Culture medium of bone marrow-derived human mesenchymal stem cells effects lymphatic endothelial cells and tumor lymph vessel formation
    Article Snippet: The human gastric carcinoma SGC-7901 and HGC-27 cell lines were purchased from the Chinese Academy of Sciences Type Culture Collection Committee cell bank (Beijing, China). .. The cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco Life Technologies, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS; Gibco) at 37°C in 5% CO2 . .. The hMSCs were cultured to ~70% confluency, and the 5 ml of medium was refreshed prior to the cells being incubated for an additional 48 h. In total, 0.22 μm filter-sterilized supernatant was collected and designated as hBM-MSC-CM.

    Article Title: Optimization of culture conditions for porcine corneal endothelial cells
    Article Snippet: Porcine eyes were obtained from a local slaughterhouse within 24 h after death. .. Corneas were dissected out of the globes using a curved scissors (Storz, St. Louis, MO) and washed several times in Dulbecco's Modified Eagle's Medium (DMEM; Invitrogen, Burlington, Ontario, Canada) containing 100 UI/ml penicillin (Sigma, Oakville, Ontario, Canada) and 25 μg/ml gentamicin (Schering Canada, Pointe-Claire, Québec, Canada). .. Corneas were incubated 30 min at 37 °C in 2.5 mg/ml dispase (Roche, Laval, Québec, Canada).

    Article Title: Cytotoxic and Antiproliferative Effect of Tepary Bean Lectins on C33-A, MCF-7, SKNSH, and SW480 Cell Lines
    Article Snippet: The cytotoxicity assay was conducted using the following four human malignant cells lines: MCF-7 (human breast adenocarcinoma); SKNSH (human bone marrow neuroblastoma); SW480 (human Caucasian colon adenocarcinoma), and C33-A (human epithelial cervical carcinoma), which were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA). .. The cells were grown in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Grand Island, NY, USA) with 10% Fetal bovine serum (FBS) (Gibco) in a CO2 water-jacketed incubator (Nuaire,Plymouth, MN, USA) at 37 °C in a humidified atmosphere of 5% CO2 and 95% air. .. The tetrazolium dye colorimetric test (3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide (MTT) was used to determine the viability of the cells lines.

    Article Title: Various Wavelengths of Light-Emitting Diode Light Regulate the Proliferation of Human Dermal Papilla Cells and Hair Follicles via Wnt/β-Catenin and the Extracellular Signal-Regulated Kinase Pathways
    Article Snippet: hDPCs (Promocell; GmbH, Heidlberg, Germany) were cultured as previously described . .. The DPCs (2×104 cells per well) were seeded into 24-well culture plates and then irradiated with LED in 10% fetal bovine serum (Gibco BRL; Life Technology, Karlsruhe, Germany), 1% penicillin/streptomycin (Gibco BRL) containing Dulbecco's modified Eagle medium (DMEM) for 48 hours. .. DPCs were used passage from 3 to 4 for the experiments.

    Article Title: Laminarin-induced apoptosis in human colon cancer LoVo cells
    Article Snippet: In addition, this study may increase the development and application of laminarin for colon cancer treatment. .. The following reagents were used: Laminarin and paraformaldehyde (Sigma-Aldrich, St. Louis, MO, USA); hydroxycamptothecin (HCPT, Harbin Shengtai Pharmaceutical Co., Ltd., Harbin, China); Dulbecco’s modified Eagle’s medium (DMEM)/F12 culture medium (Thermo Fisher Scientific Inc., Waltham, MA, USA); fetal bovine serum (FBS; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China); pancreatin (Gibco-BRL, Rockville, MD, USA); Hoechst 33258 (Sigma-Aldrich); rabbit anti-human β-actin polyclonal antibody, DR4, DR5, TNF-related apoptosis-inducing ligand (TRAIL), Fas-associated protein with death domain (FADD) and Bid (Biosynthesis Biotechnology Co., Ltd., Beijing, China); mouse anti-human Bcl-2, rabbit anti-human Bax and fluorescein isothiocyanate (FITC)-goat anti-mouse polyclonal antibodies (Boster Biological Technology Co., Ltd., Wuhan, China); mouse anti-human caspase-8 and -3, alkaline phosphatase goat anti-rabbit polyclonal antibodies, as well as caspase-8, -3, -6 and -7 activity assay kits, sodium dodecyl sulphate sample buffer, polyacrylamide gel, nitrocellulose membranes, 5-bromo-4-chloro-3-indolyl-phosphate (BCIP)/nitroblue tetrazolium (NBT) alkaline phosphatase color development kit, Triton X-100, bovine serum albumin and phosphate-buffered saline (PBS; Beyotime Institute of Biotechnology, Haimen, China); detergent-compatible protein assay kit (Bio-Rad, Hercules, CA, USA). .. The CKX41 fluorescence inverted microscope was purchased from Olympus (Tokyo, Japan), and the Mini-Protean Tetra and Mini Trans-Blot electrophoresis systems, Gel Doc XR imaging system and Model 680 microplate reader were purchased from Bio-Rad.

    Article Title: Effect of fibroblast growth factor 9 on the osteogenic differentiation of bone marrow stromal stem cells and dental pulp stem cells
    Article Snippet: Following cleaning of the teeth, the pulp chambers were accessed by cutting the cementoenamel junction with a sterile fissure dental bur. .. Following exposure of the pulp, the pulp tissue was removed in fragments of ~0.5 mm, which were then placed onto a 6-cm culture dish containing Dulbecco’s modified Eagle’s medium (DMEM; Gibco-BRL, Grand Island, NY, USA) supplemented with 20% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) and antibiotics (Gibco-BRL) prior to incubation at 37°C in 5% CO2 . .. At confluence, the outgrown cells were transferred onto a 10-cm dish and then continuously passaged for further experiments.

    Article Title: Transforming growth factor-β decreases side population cells in hepatocellular carcinoma in vitro
    Article Snippet: Huh-7 and Huh-Bat [Huh-7 transfected with human Na+ -taurocholate co-transporting polypeptide (bile acid transporter) cDNA] cell lines were obtained from the Korean Cell Line Bank (Seoul, Korea). .. Cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Inc.). .. For adherent cultures, 5×105 cells were seeded onto tissue culture dishes (BD Biosciences, San Jose, CA, USA).

    Article Title: Diet-induced alteration of fatty acid synthase in prostate cancer progression
    Article Snippet: Human PCa LNCaP cells were purchased from the American Type Culture Collection (Manassas, VA, USA) and the PCa C4-2 cells were kindly provided by Dr Leland WK Chung of Emery University. .. The cells were maintained in RPMI 1640 medium or Dulbecco's modified Eagle's medium (Invitrogen, Carlsbad, CA, USA) containing 10% fetal bovine serum and 1% penicillin–streptomycin. .. PI3K inhibitor (LY294002), MAPK inhibitor (U0126) and AMPK activator (AICAR) were purchased from Cell Signaling Technology (Boston, MA, USA).

    Article Title: Inhibitory Effects of Enalaprilat on Rat Cardiac Fibroblast Proliferation via ROS/P38MAPK/TGF-β1 Signaling Pathway
    Article Snippet: The dishes were gently rinsed three times to remove the remaining cardiomyocyte. .. The culture medium for cardiac fibroblasts was changed for Dulbecco’s Modified Eagle Medium (DMEM; Gibco BRL) supplemented with 20% FCS and 25 μg/mL gentamycin. .. Purity of cardiac fibroblast culture ( > 97%) was assessed with repeated differential plating, microscopic evaluation and immunostaining.

    Article Title: MMP‐2 and MMP‐14 Silencing Inhibits VEGFR2 Cleavage and Induces the Differentiation of Porcine Adipose‐Derived Mesenchymal Stem Cells to Endothelial Cells
    Article Snippet: MSCs were harvested from porcine abdominal fat with slight differences in the isolation procedure from previous published studies , , , . .. The adipose tissue from the abdominal fat was collected from the slaughterhouse and transferred to the laboratory in sterile conditions in Dulbecco's modified Eagle medium (DMEM; Invitrogen, Grand Island, NY, http://www.invitrogen.com ) with antibiotics 100 mg/ml penicillin (Sigma‐Aldrich, St. Louis, MO, http://www.sigmaaldrich.com ), and 100 mg/ml streptomycin (Sigma‐Aldrich). .. The transferred adipose tissue was washed extensively with excess amount of phosphate‐buffered saline (PBS), and minced into 4–8 mm pieces with sterile scissors.

    Article Title: Effects of bradykinin on TGF-β1-induced epithelial-mesenchymal transition in ARPE-19 cells
    Article Snippet: Human retinal pigment epithelial cells (ARPE-19) were obtained from Cell Biosciences Pty, Ltd. (Heidelberg, Australia). .. ARPE-19 cells were cultured in Dulbecco's modified Eagle's medium (DMEM)/H (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.). .. Cells were incubated at 37°C in a humidified incubator containing 5% CO2 .

    Article Title: The Inhibitory Effects of Forsythia Koreana Extracts on the Metastatic Ability of Breast Cancer Cells and Bone Resorption by Osteoclasts
    Article Snippet: The extracts were dissolved with dimethyl sulfoxide (DMSO). .. Dulbecco’s modified Eagle medium (DMEM), FBS, Dulbecco’s PBS, α-minimum essential medium (α-MEM), and antibiotics were purchased from Gibco BRL (Grand Island, NY, USA). .. Recombinant mouse soluble RANKL (sRANKL) and macrophage-colony stimulating factor (M-CSF) were obtained from R & D Systems (Minneapolis, MN, USA).

    Article Title: Sonic hedgehog (SHH) signaling improves the angiogenic potential of Wharton’s jelly-derived mesenchymal stem cells (WJ-MSC)
    Article Snippet: WJ-MSC were isolated and characterized as previously described with minor modifications [ ]. .. Briefly, UCs were stored and transported in Dulbecco’s modified Eagle’s medium (DMEM; Thermo Scientific) from maternity facilities to our laboratory and were processed within 24 h postdelivery. .. Tissues were cut into 2 mm2 pieces and blood vessels were discarded.

    Article Title: Oseltamivir phosphate monotherapy ablates tumor neovascularization, growth, and metastasis in mouse model of human triple-negative breast adenocarcinoma
    Article Snippet: MCF-7 is a non-triple negative human breast adenocarcinoma cell line. .. The cells were grown in culture media containing 1× Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Rockville, MD, USA), conditioned medium, supplemented with 10% fetal calf serum (FCS; HyClone, Logan, UT, USA), and 5 μg/mL plasmocin™ (InvivoGen, San Diego, CA, USA) in a 5% CO2 incubator at 37°C. .. MCF-7 and MDA-MB-231 cell lines resistant against 5 μM and 10 μM tamoxifen were established in culture to gradual increases in concentrations of the indicated drugs in 1× DMEM conditioned medium.

    Western Blot:

    Article Title: Pharmacological disruption of insulin-like growth factor 1 binding to IGF-binding proteins restores anabolic responses in human osteoarthritic chondrocytes
    Article Snippet: Paragraph title: Western ligand blot analysis of IGFBPs ... Explants were dissected out from cartilage, washed extensively with Dulbecco's phosphate-buffered saline without calcium and magnesium (DPBS, Gibco), and incubated directly in serum-free Ham F12 medium without prior incubation in FCS-containing medium.

    Cytotoxicity Assay:

    Article Title: Cytotoxic and Antiproliferative Effect of Tepary Bean Lectins on C33-A, MCF-7, SKNSH, and SW480 Cell Lines
    Article Snippet: The cytotoxicity assay was conducted using the following four human malignant cells lines: MCF-7 (human breast adenocarcinoma); SKNSH (human bone marrow neuroblastoma); SW480 (human Caucasian colon adenocarcinoma), and C33-A (human epithelial cervical carcinoma), which were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA). .. The cells were grown in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Grand Island, NY, USA) with 10% Fetal bovine serum (FBS) (Gibco) in a CO2 water-jacketed incubator (Nuaire,Plymouth, MN, USA) at 37 °C in a humidified atmosphere of 5% CO2 and 95% air.

    Derivative Assay:

    Article Title: Oseltamivir phosphate monotherapy ablates tumor neovascularization, growth, and metastasis in mouse model of human triple-negative breast adenocarcinoma
    Article Snippet: MCF-7 (ATCC® HTB-22™) and MDA-MB-231 (ATCC® HTB-26™) are adherent epithelial adenocarcinomas obtained from the mammary gland, breast, which are derived from the metastatic pleural effusion site. .. The cells were grown in culture media containing 1× Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Rockville, MD, USA), conditioned medium, supplemented with 10% fetal calf serum (FCS; HyClone, Logan, UT, USA), and 5 μg/mL plasmocin™ (InvivoGen, San Diego, CA, USA) in a 5% CO2 incubator at 37°C.

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    Thermo Fisher dulbecco
    Effect of laminarin on apoptosis-related protein expression in LoVo cells. The LoVo cells were treated with low (400 μg/ml), middle (800 μg/ml) and high (1,600 μg/ml) concentrations of lamarin, as well as <t>Dulbecco’s</t> modified Eagle’s medium/F12 culture medium as a control and HCPT as a negative control. DR, death receptor; TRAIL, TNF-related apoptosis-inducing ligand; FADD, FAS-associated protein with death domain; Casp, caspase; HCPT, hydroxycamptothecin.
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    Effect of laminarin on apoptosis-related protein expression in LoVo cells. The LoVo cells were treated with low (400 μg/ml), middle (800 μg/ml) and high (1,600 μg/ml) concentrations of lamarin, as well as Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and HCPT as a negative control. DR, death receptor; TRAIL, TNF-related apoptosis-inducing ligand; FADD, FAS-associated protein with death domain; Casp, caspase; HCPT, hydroxycamptothecin.

    Journal: Oncology Letters

    Article Title: Laminarin-induced apoptosis in human colon cancer LoVo cells

    doi: 10.3892/ol.2014.1952

    Figure Lengend Snippet: Effect of laminarin on apoptosis-related protein expression in LoVo cells. The LoVo cells were treated with low (400 μg/ml), middle (800 μg/ml) and high (1,600 μg/ml) concentrations of lamarin, as well as Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and HCPT as a negative control. DR, death receptor; TRAIL, TNF-related apoptosis-inducing ligand; FADD, FAS-associated protein with death domain; Casp, caspase; HCPT, hydroxycamptothecin.

    Article Snippet: The following reagents were used: Laminarin and paraformaldehyde (Sigma-Aldrich, St. Louis, MO, USA); hydroxycamptothecin (HCPT, Harbin Shengtai Pharmaceutical Co., Ltd., Harbin, China); Dulbecco’s modified Eagle’s medium (DMEM)/F12 culture medium (Thermo Fisher Scientific Inc., Waltham, MA, USA); fetal bovine serum (FBS; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China); pancreatin (Gibco-BRL, Rockville, MD, USA); Hoechst 33258 (Sigma-Aldrich); rabbit anti-human β-actin polyclonal antibody, DR4, DR5, TNF-related apoptosis-inducing ligand (TRAIL), Fas-associated protein with death domain (FADD) and Bid (Biosynthesis Biotechnology Co., Ltd., Beijing, China); mouse anti-human Bcl-2, rabbit anti-human Bax and fluorescein isothiocyanate (FITC)-goat anti-mouse polyclonal antibodies (Boster Biological Technology Co., Ltd., Wuhan, China); mouse anti-human caspase-8 and -3, alkaline phosphatase goat anti-rabbit polyclonal antibodies, as well as caspase-8, -3, -6 and -7 activity assay kits, sodium dodecyl sulphate sample buffer, polyacrylamide gel, nitrocellulose membranes, 5-bromo-4-chloro-3-indolyl-phosphate (BCIP)/nitroblue tetrazolium (NBT) alkaline phosphatase color development kit, Triton X-100, bovine serum albumin and phosphate-buffered saline (PBS; Beyotime Institute of Biotechnology, Haimen, China); detergent-compatible protein assay kit (Bio-Rad, Hercules, CA, USA).

    Techniques: Expressing, Modification, Negative Control

    Effect of laminarin on Bid and tBid expression in LoVo cells. The LoVo cells were treated with low (400 μg/ml), middle (800 μg/ml) and high (1,600 μg/ml) concentrations of lamarin, as well as Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and HCPT as a negative control. HCPT, hydroxycamptothecin.

    Journal: Oncology Letters

    Article Title: Laminarin-induced apoptosis in human colon cancer LoVo cells

    doi: 10.3892/ol.2014.1952

    Figure Lengend Snippet: Effect of laminarin on Bid and tBid expression in LoVo cells. The LoVo cells were treated with low (400 μg/ml), middle (800 μg/ml) and high (1,600 μg/ml) concentrations of lamarin, as well as Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and HCPT as a negative control. HCPT, hydroxycamptothecin.

    Article Snippet: The following reagents were used: Laminarin and paraformaldehyde (Sigma-Aldrich, St. Louis, MO, USA); hydroxycamptothecin (HCPT, Harbin Shengtai Pharmaceutical Co., Ltd., Harbin, China); Dulbecco’s modified Eagle’s medium (DMEM)/F12 culture medium (Thermo Fisher Scientific Inc., Waltham, MA, USA); fetal bovine serum (FBS; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China); pancreatin (Gibco-BRL, Rockville, MD, USA); Hoechst 33258 (Sigma-Aldrich); rabbit anti-human β-actin polyclonal antibody, DR4, DR5, TNF-related apoptosis-inducing ligand (TRAIL), Fas-associated protein with death domain (FADD) and Bid (Biosynthesis Biotechnology Co., Ltd., Beijing, China); mouse anti-human Bcl-2, rabbit anti-human Bax and fluorescein isothiocyanate (FITC)-goat anti-mouse polyclonal antibodies (Boster Biological Technology Co., Ltd., Wuhan, China); mouse anti-human caspase-8 and -3, alkaline phosphatase goat anti-rabbit polyclonal antibodies, as well as caspase-8, -3, -6 and -7 activity assay kits, sodium dodecyl sulphate sample buffer, polyacrylamide gel, nitrocellulose membranes, 5-bromo-4-chloro-3-indolyl-phosphate (BCIP)/nitroblue tetrazolium (NBT) alkaline phosphatase color development kit, Triton X-100, bovine serum albumin and phosphate-buffered saline (PBS; Beyotime Institute of Biotechnology, Haimen, China); detergent-compatible protein assay kit (Bio-Rad, Hercules, CA, USA).

    Techniques: Expressing, Modification, Negative Control

    Effect of laminarin on LoVo cell morphology. LoVo cells were treated with (A) Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and (B) hydroxycamptothecin (5 μg/ml) as a negative control, as well as (C) low (400 μg/ml), (D) middle (800 μg/ml) and (E) high (1600 μg/ml) concentrations of laminarin. Arrows indicate apoptotic bodies.

    Journal: Oncology Letters

    Article Title: Laminarin-induced apoptosis in human colon cancer LoVo cells

    doi: 10.3892/ol.2014.1952

    Figure Lengend Snippet: Effect of laminarin on LoVo cell morphology. LoVo cells were treated with (A) Dulbecco’s modified Eagle’s medium/F12 culture medium as a control and (B) hydroxycamptothecin (5 μg/ml) as a negative control, as well as (C) low (400 μg/ml), (D) middle (800 μg/ml) and (E) high (1600 μg/ml) concentrations of laminarin. Arrows indicate apoptotic bodies.

    Article Snippet: The following reagents were used: Laminarin and paraformaldehyde (Sigma-Aldrich, St. Louis, MO, USA); hydroxycamptothecin (HCPT, Harbin Shengtai Pharmaceutical Co., Ltd., Harbin, China); Dulbecco’s modified Eagle’s medium (DMEM)/F12 culture medium (Thermo Fisher Scientific Inc., Waltham, MA, USA); fetal bovine serum (FBS; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd., Hangzhou, China); pancreatin (Gibco-BRL, Rockville, MD, USA); Hoechst 33258 (Sigma-Aldrich); rabbit anti-human β-actin polyclonal antibody, DR4, DR5, TNF-related apoptosis-inducing ligand (TRAIL), Fas-associated protein with death domain (FADD) and Bid (Biosynthesis Biotechnology Co., Ltd., Beijing, China); mouse anti-human Bcl-2, rabbit anti-human Bax and fluorescein isothiocyanate (FITC)-goat anti-mouse polyclonal antibodies (Boster Biological Technology Co., Ltd., Wuhan, China); mouse anti-human caspase-8 and -3, alkaline phosphatase goat anti-rabbit polyclonal antibodies, as well as caspase-8, -3, -6 and -7 activity assay kits, sodium dodecyl sulphate sample buffer, polyacrylamide gel, nitrocellulose membranes, 5-bromo-4-chloro-3-indolyl-phosphate (BCIP)/nitroblue tetrazolium (NBT) alkaline phosphatase color development kit, Triton X-100, bovine serum albumin and phosphate-buffered saline (PBS; Beyotime Institute of Biotechnology, Haimen, China); detergent-compatible protein assay kit (Bio-Rad, Hercules, CA, USA).

    Techniques: Modification, Negative Control

    Cell cycle analysis of Huh-7 and Huh-Bat cells following SB431542 and TGF-β treatment. Huh-7 or Huh-Bat cells (5×105 ) were seeded onto Dulbecco's modified Eagle's medium containing 10% fetal bovine serum. Cells were incubated for 72 h with dimethyl sulfoxide (control), 1 µM SB431542, 1 ng/ml TGF-β or 1 µM SB431542 plus 1 ng/ml TGF-β. Cells were stained with propidium iodide and analyzed by flow cytometry. (A) Huh-7 cells. (B) Huh-Bat cells. TGF-β, transforming growth factor-β; CTL, control.

    Journal:

    Article Title: Transforming growth factor-β decreases side population cells in hepatocellular carcinoma in vitro

    doi: 10.3892/ol.2018.8441

    Figure Lengend Snippet: Cell cycle analysis of Huh-7 and Huh-Bat cells following SB431542 and TGF-β treatment. Huh-7 or Huh-Bat cells (5×105 ) were seeded onto Dulbecco's modified Eagle's medium containing 10% fetal bovine serum. Cells were incubated for 72 h with dimethyl sulfoxide (control), 1 µM SB431542, 1 ng/ml TGF-β or 1 µM SB431542 plus 1 ng/ml TGF-β. Cells were stained with propidium iodide and analyzed by flow cytometry. (A) Huh-7 cells. (B) Huh-Bat cells. TGF-β, transforming growth factor-β; CTL, control.

    Article Snippet: Cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Inc.).

    Techniques: Cell Cycle Assay, Modification, Incubation, Staining, Flow Cytometry, Cytometry, CTL Assay

    Cell survival of Huh-7 and Huh-Bat cells following SB431542 and TGF-β treatment. Cells (5×105 ) were seeded onto plates in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum. Cultures were incubated for 72 h with dimethyl sulfoxide (control), 1 µM SB431542, 1 ng/ml TGF-β or 1 µM SB431542 plus 1 ng/ml TGF-β. Cell survival was assessed by cell counting using trypan blue dye exclusion. (A) Huh-7 cells. (B) Huh-Bat cells. The relative cell survival rate is shown as survival percentage vs. control cells. Values are presented as the mean ± standard error from at least three independent experiments. *P≤0.05; **P≤0.01. TGF-β, transforming growth factor-β; CTL, control.

    Journal:

    Article Title: Transforming growth factor-β decreases side population cells in hepatocellular carcinoma in vitro

    doi: 10.3892/ol.2018.8441

    Figure Lengend Snippet: Cell survival of Huh-7 and Huh-Bat cells following SB431542 and TGF-β treatment. Cells (5×105 ) were seeded onto plates in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum. Cultures were incubated for 72 h with dimethyl sulfoxide (control), 1 µM SB431542, 1 ng/ml TGF-β or 1 µM SB431542 plus 1 ng/ml TGF-β. Cell survival was assessed by cell counting using trypan blue dye exclusion. (A) Huh-7 cells. (B) Huh-Bat cells. The relative cell survival rate is shown as survival percentage vs. control cells. Values are presented as the mean ± standard error from at least three independent experiments. *P≤0.05; **P≤0.01. TGF-β, transforming growth factor-β; CTL, control.

    Article Snippet: Cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Inc.).

    Techniques: Modification, Incubation, Cell Counting, CTL Assay

    SHH pathway is active in WJ-MSC in vivo and enhances their pro-angiogenic properties. In vitro pretreated Wharton’s jelly-derived mesenchymal stem cells ( WJ-MSC ) (+ N-Shh or Cyc) were applied on top of the chorioallantoic membrane ( CAM ) of chicken embryos. Treatment was repeated after 48 h in order to maintain the effect on SHH pathway modulation in WJ-MSC. The angiogenic response was evaluated after 96 h. Recombinant fibroblast growth factor 2 ( FGF2 ), a potent angiogenic stimulator, was used as a positive control ( a ), and ( b ) Dulbecco’s modified Eagle’s medium ( DMEM ) was used as negative control (WJ-MSC vehicle). c WJ-MSC seeded in Integra Matrix ( IM ). WJ-MSC seeded in Integra Matrix plus N-Shh ( d ) and Cyc ( e ). f Assay quantification. * P

    Journal: Stem Cell Research & Therapy

    Article Title: Sonic hedgehog (SHH) signaling improves the angiogenic potential of Wharton’s jelly-derived mesenchymal stem cells (WJ-MSC)

    doi: 10.1186/s13287-017-0653-8

    Figure Lengend Snippet: SHH pathway is active in WJ-MSC in vivo and enhances their pro-angiogenic properties. In vitro pretreated Wharton’s jelly-derived mesenchymal stem cells ( WJ-MSC ) (+ N-Shh or Cyc) were applied on top of the chorioallantoic membrane ( CAM ) of chicken embryos. Treatment was repeated after 48 h in order to maintain the effect on SHH pathway modulation in WJ-MSC. The angiogenic response was evaluated after 96 h. Recombinant fibroblast growth factor 2 ( FGF2 ), a potent angiogenic stimulator, was used as a positive control ( a ), and ( b ) Dulbecco’s modified Eagle’s medium ( DMEM ) was used as negative control (WJ-MSC vehicle). c WJ-MSC seeded in Integra Matrix ( IM ). WJ-MSC seeded in Integra Matrix plus N-Shh ( d ) and Cyc ( e ). f Assay quantification. * P

    Article Snippet: Briefly, UCs were stored and transported in Dulbecco’s modified Eagle’s medium (DMEM; Thermo Scientific) from maternity facilities to our laboratory and were processed within 24 h postdelivery.

    Techniques: In Vivo, In Vitro, Derivative Assay, Chick Chorioallantoic Membrane Assay, Recombinant, Positive Control, Modification, Negative Control

    Bone marrow mesenchymal stem cell viability was assessed by MTT assay after 24 h of exposure to low-glucose DMEM supplemented with 1, 10, 50, 100, 200, 500 and 1,000 µM PVP-I. DMEM represents the PVP-I-free control group. DMEM, Dulbecco's modified Eagle's medium; PVP-I, polyvinylpyrrolidone-iodine.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Effects of polyvinylpyrrolidone-iodine on tendon-bone healing in a rabbit extra-articular model

    doi: 10.3892/etm.2017.4359

    Figure Lengend Snippet: Bone marrow mesenchymal stem cell viability was assessed by MTT assay after 24 h of exposure to low-glucose DMEM supplemented with 1, 10, 50, 100, 200, 500 and 1,000 µM PVP-I. DMEM represents the PVP-I-free control group. DMEM, Dulbecco's modified Eagle's medium; PVP-I, polyvinylpyrrolidone-iodine.

    Article Snippet: In brief, 2 ml bone marrow was obtained, mixed with 2 ml low-glucose Dulbecco's modified Eagle's medium (DMEM; Thermo Fisher Scientific, Inc., Waltham, MA, USA) and centrifuged at 1,200 × g for 5 min.

    Techniques: MTT Assay, Modification