penicillin streptomycin solution  (Thermo Fisher)


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    Name:
    Shandon 10000 Autopsy Saw
    Description:
    Cut safely and reduce operator fatigue with Thermo Scientific Shandon 10000 Autopsy Saw
    Catalog Number:
    10000
    Price:
    None
    Category:
    Instruments and Equipment
    Applications:
    Anatomical Pathology|Clinical
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    Thermo Fisher penicillin streptomycin solution
    Cut safely and reduce operator fatigue with Thermo Scientific Shandon 10000 Autopsy Saw
    https://www.bioz.com/result/penicillin streptomycin solution/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    penicillin streptomycin solution - by Bioz Stars, 2021-03
    99/100 stars

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    Cell Culture:

    Article Title: Bioprinting of stem cell expansion lattices
    Article Snippet: NPCs were cultured following a previously described protocol [ ]. .. Briefly, NPCs were cultured in Stemness Maintenance Medium (Neurobasal-A, 2% B27 Supplement with Vitamin A (Gibco), GlutaMAX (Gibco), 20 ng ml−1 FGF-2 (PeproTech), 20 ng ml−1 EGF (PeproTech), and 1% Pen/Strep (Gibco)) on tissue culture plastic coated with 10 μg mL−1 polyornithine (Sigma-Aldrich) and 5 mg mL−1 laminin (Gibco) at a seeding density of 10,000 NPCs cm−2 . ..

    Mutagenesis:

    Article Title: Specific bone cells produce DLL4 to generate thymus-seeding progenitors from bone marrow
    Article Snippet: .. 35,000 or 10,000 Ly6D- CLPs (Lin− Sca+ cKit+ IL7R+ Thy1.2− Ly6D− ) were sorted from OcnCre;iDTR control and mutant donors (CD45.2-Pacific Blue), labeled with 10 µM CFDA-SE (Invitrogen) and 1 µg/ml VivoTag680 (VT680, VisEn Medical), respectively, mixed in a 1:1 ratio, and transplanted into each congenic SJL recipient (CD45.1-PE) by retro-orbital injection. .. The same number of progenitor cells sorted from CCR7−/− mice served as another control to compete with OcnCre;iDTR mutant donors.

    Labeling:

    Article Title: Specific bone cells produce DLL4 to generate thymus-seeding progenitors from bone marrow
    Article Snippet: .. 35,000 or 10,000 Ly6D- CLPs (Lin− Sca+ cKit+ IL7R+ Thy1.2− Ly6D− ) were sorted from OcnCre;iDTR control and mutant donors (CD45.2-Pacific Blue), labeled with 10 µM CFDA-SE (Invitrogen) and 1 µg/ml VivoTag680 (VT680, VisEn Medical), respectively, mixed in a 1:1 ratio, and transplanted into each congenic SJL recipient (CD45.1-PE) by retro-orbital injection. .. The same number of progenitor cells sorted from CCR7−/− mice served as another control to compete with OcnCre;iDTR mutant donors.

    Injection:

    Article Title: Specific bone cells produce DLL4 to generate thymus-seeding progenitors from bone marrow
    Article Snippet: .. 35,000 or 10,000 Ly6D- CLPs (Lin− Sca+ cKit+ IL7R+ Thy1.2− Ly6D− ) were sorted from OcnCre;iDTR control and mutant donors (CD45.2-Pacific Blue), labeled with 10 µM CFDA-SE (Invitrogen) and 1 µg/ml VivoTag680 (VT680, VisEn Medical), respectively, mixed in a 1:1 ratio, and transplanted into each congenic SJL recipient (CD45.1-PE) by retro-orbital injection. .. The same number of progenitor cells sorted from CCR7−/− mice served as another control to compete with OcnCre;iDTR mutant donors.

    Transfection:

    Article Title: Novel lincRNA SLINKY is a prognostic biomarker in kidney cancer
    Article Snippet: .. siRNA transfections and proliferation assays ccRCC cells were seeded (10,000 cells per well) in 6-well plates and transfected using Lipofectamine 2000 (Life Technologies). .. Two independent small interfering (si)RNAs targeting the first exon of SLINKY (On-TARGETplus, Dharmacon) were transfected at a final siRNA concentration of 50nM for 16 hrs (custom-designed siRNA sequences available in ).

    Multiple Displacement Amplification:

    Article Title: Molecular Imaging of the Translocator Protein (TSPO) in a Pre-Clinical Model of Breast Cancer
    Article Snippet: A Nikon Eclipse TE2000-U fluorescence microscope equipped with a mercury lamp, indocyanine green filter set and a Hamamatsu ORCA II BT 512 camera controlled by Metamorph v6.1 (Molecular Devices Corporation; Downingtown, PA) was used for imaging. .. MDA-MB-231 cells were plated at 10,000 cells per well in parafilm-wrapped 96 MicroWell™ Nunclon™Δ Optical Bottom Plates (Nalge Nunc International; Rochester, NY) and incubated under standard culture conditions for approximately 48 h. Immediately prior to experimentation, the cells were washed once with 37°C FBS-free medium to remove any dead cells and serum. .. The cells were then divided into four populations and evaluated in triplicate: (1) cells incubated with increasing concentrations of NIR-conPK11195 (1, 4, 7, 10, 40, 70, 100, 400, 700 nM, 1 μM) in FBS-free medium (100 μL volume), (2) cells simultaneously incubated with the same concentrations of NIR-conPK11195 and 100 µM PK 11195, (3) cells incubated with the same concentrations of free NIR dye, and (4) undosed cells as blanks.

    Incubation:

    Article Title: Molecular Imaging of the Translocator Protein (TSPO) in a Pre-Clinical Model of Breast Cancer
    Article Snippet: A Nikon Eclipse TE2000-U fluorescence microscope equipped with a mercury lamp, indocyanine green filter set and a Hamamatsu ORCA II BT 512 camera controlled by Metamorph v6.1 (Molecular Devices Corporation; Downingtown, PA) was used for imaging. .. MDA-MB-231 cells were plated at 10,000 cells per well in parafilm-wrapped 96 MicroWell™ Nunclon™Δ Optical Bottom Plates (Nalge Nunc International; Rochester, NY) and incubated under standard culture conditions for approximately 48 h. Immediately prior to experimentation, the cells were washed once with 37°C FBS-free medium to remove any dead cells and serum. .. The cells were then divided into four populations and evaluated in triplicate: (1) cells incubated with increasing concentrations of NIR-conPK11195 (1, 4, 7, 10, 40, 70, 100, 400, 700 nM, 1 μM) in FBS-free medium (100 μL volume), (2) cells simultaneously incubated with the same concentrations of NIR-conPK11195 and 100 µM PK 11195, (3) cells incubated with the same concentrations of free NIR dye, and (4) undosed cells as blanks.

    Real-time Polymerase Chain Reaction:

    Article Title: Seven Novel Probe Systems for Real-Time PCR Provide Absolute Single-Base Discrimination, Higher Signaling, and Generic Components
    Article Snippet: In Universal probe assays, the concentration of linker primer used in the first-step reaction was 20 nmol/L, and Universal probe and antiprobe concentrations were 200 and 400 nmol/L, respectively. .. We typically used 10 ng of viral cDNA or 1000 to 10,000 copies of ultramer template per reaction. qPCR reactions were performed in 25-μL volumes containing 12.5 μL of 2X HotStart-IT Probe qPCR Master Mix (Affymetrix, Inc., Santa Clara, CA), the indicated template, primer, probe, and/or antiprobe concentrations, and additional MgCl2 and dNTPs (added at final concentrations of 5 and 0.1 mmol/L, respectively). .. Typical thermal cycling conditions are 95°C, 5 minutes, and then 40 cycles of either two-step PCR (denaturation at 95°C for 15 seconds, and annealing/extension at 58°C for 30 to 60 seconds), or three-step PCR (denaturation at 95°C, 15 seconds; annealing at 58°C, 45 seconds; and extension at 72°C, 45 seconds).

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  • 98
    Thermo Fisher lpds
    ATGL expression in macrophages and foam cells. Mouse peritoneal macrophages, human THP-1, and human monocyte-derived macrophages ( HMDM ) were cultivated in <t>DMEM,</t> 10% <t>LPDS</t> in the absence (control) or presence of 100 μg of acLDL/ml. Total RNA was isolated and reverse-transcribed, and mRNA expression of ATGL ( A and C ) and hormone-sensitive lipase ( HSL ) was determined by real time PCR, including murine hypoxanthine-guanine phosphoribosyltransferase or human ( hu ) porphobilinogen deaminase normalization ( A ). Untreated macrophages were arbitrarily set to 1. Data are expressed as mean values ( n = 3) ± S.E. of triplicate repeats. ***, p ≤ 0.001. B , cell extracts of macrophages, foam cells (40 μg per lane), and white adipose tissue ( WAT ) (10 and 40 μg per lane) were resolved by SDS-PAGE. Protein expression of ATGL and hormone-sensitive lipase were analyzed by Western blotting relative to the expression of β-actin. STD , standard.
    Lpds, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lpds/product/Thermo Fisher
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    ATGL expression in macrophages and foam cells. Mouse peritoneal macrophages, human THP-1, and human monocyte-derived macrophages ( HMDM ) were cultivated in DMEM, 10% LPDS in the absence (control) or presence of 100 μg of acLDL/ml. Total RNA was isolated and reverse-transcribed, and mRNA expression of ATGL ( A and C ) and hormone-sensitive lipase ( HSL ) was determined by real time PCR, including murine hypoxanthine-guanine phosphoribosyltransferase or human ( hu ) porphobilinogen deaminase normalization ( A ). Untreated macrophages were arbitrarily set to 1. Data are expressed as mean values ( n = 3) ± S.E. of triplicate repeats. ***, p ≤ 0.001. B , cell extracts of macrophages, foam cells (40 μg per lane), and white adipose tissue ( WAT ) (10 and 40 μg per lane) were resolved by SDS-PAGE. Protein expression of ATGL and hormone-sensitive lipase were analyzed by Western blotting relative to the expression of β-actin. STD , standard.

    Journal: The Journal of Biological Chemistry

    Article Title: Efficient Phagocytosis Requires Triacylglycerol Hydrolysis by Adipose Triglyceride Lipase *

    doi: 10.1074/jbc.M110.107854

    Figure Lengend Snippet: ATGL expression in macrophages and foam cells. Mouse peritoneal macrophages, human THP-1, and human monocyte-derived macrophages ( HMDM ) were cultivated in DMEM, 10% LPDS in the absence (control) or presence of 100 μg of acLDL/ml. Total RNA was isolated and reverse-transcribed, and mRNA expression of ATGL ( A and C ) and hormone-sensitive lipase ( HSL ) was determined by real time PCR, including murine hypoxanthine-guanine phosphoribosyltransferase or human ( hu ) porphobilinogen deaminase normalization ( A ). Untreated macrophages were arbitrarily set to 1. Data are expressed as mean values ( n = 3) ± S.E. of triplicate repeats. ***, p ≤ 0.001. B , cell extracts of macrophages, foam cells (40 μg per lane), and white adipose tissue ( WAT ) (10 and 40 μg per lane) were resolved by SDS-PAGE. Protein expression of ATGL and hormone-sensitive lipase were analyzed by Western blotting relative to the expression of β-actin. STD , standard.

    Article Snippet: Macrophages were cultured in DMEM, 10% LPDS (Invitrogen) supplemented with 1% penicillin, 1% streptomycin for 2 h. Thereafter, cells were washed twice with PBS, and if not stated otherwise, the adherent macrophages were cultured in high glucose (25 mm ) DMEM containing 4 mm glutamine, 1 mm pyruvate, and 10% LPDS.

    Techniques: Expressing, Derivative Assay, Isolation, Real-time Polymerase Chain Reaction, SDS Page, Western Blot

    mRNA levels of mitochondrial genes in Atgl −/− and WT macrophages and foam cells. MPM were cultivated in DMEM, 10% LPDS in the absence or presence of 100 μg of acLDL/ml (foam cells). Total RNA was isolated from cells and reverse-transcribed, and mRNA levels of carnitine palmitoyltransferase ( CPT )1a ( A ) and long chain acyl-CoA dehydrogenase ( LCAD ) ( B ) were determined by real time PCR, including normalization to hypoxanthine-guanine phosphoribosyltransferase ( HPRT ) levels. Untreated MPM were arbitrarily set to 1. Data are expressed as mean values ± S.E. of triplicate repeats. C , long chain acyl-CoA. D , carnitine ester concentrations were determined in macrophages of Atgl −/− and WT mice. Data are expressed as mean values ( n = 4–5) ± S.E. correlated to protein concentrations. *, p

    Journal: The Journal of Biological Chemistry

    Article Title: Efficient Phagocytosis Requires Triacylglycerol Hydrolysis by Adipose Triglyceride Lipase *

    doi: 10.1074/jbc.M110.107854

    Figure Lengend Snippet: mRNA levels of mitochondrial genes in Atgl −/− and WT macrophages and foam cells. MPM were cultivated in DMEM, 10% LPDS in the absence or presence of 100 μg of acLDL/ml (foam cells). Total RNA was isolated from cells and reverse-transcribed, and mRNA levels of carnitine palmitoyltransferase ( CPT )1a ( A ) and long chain acyl-CoA dehydrogenase ( LCAD ) ( B ) were determined by real time PCR, including normalization to hypoxanthine-guanine phosphoribosyltransferase ( HPRT ) levels. Untreated MPM were arbitrarily set to 1. Data are expressed as mean values ± S.E. of triplicate repeats. C , long chain acyl-CoA. D , carnitine ester concentrations were determined in macrophages of Atgl −/− and WT mice. Data are expressed as mean values ( n = 4–5) ± S.E. correlated to protein concentrations. *, p

    Article Snippet: Macrophages were cultured in DMEM, 10% LPDS (Invitrogen) supplemented with 1% penicillin, 1% streptomycin for 2 h. Thereafter, cells were washed twice with PBS, and if not stated otherwise, the adherent macrophages were cultured in high glucose (25 mm ) DMEM containing 4 mm glutamine, 1 mm pyruvate, and 10% LPDS.

    Techniques: Isolation, Cycling Probe Technology, Real-time Polymerase Chain Reaction, Mouse Assay

    Phagocytosis in Atgl −/− and WT macrophages. A , MPM from Atgl −/− and WT mice were cultivated in DMEM, 10% LPDS with 0, 6, and 25 m m glucose for 1 and 8 h, respectively. Phagocytosis of fluorescein-labeled E. coli particles is presented as mean values ( n = 6) ± S.E. of two independent experiments. Phagocytosis of WT cells was arbitrarily set to 100%. *, p

    Journal: The Journal of Biological Chemistry

    Article Title: Efficient Phagocytosis Requires Triacylglycerol Hydrolysis by Adipose Triglyceride Lipase *

    doi: 10.1074/jbc.M110.107854

    Figure Lengend Snippet: Phagocytosis in Atgl −/− and WT macrophages. A , MPM from Atgl −/− and WT mice were cultivated in DMEM, 10% LPDS with 0, 6, and 25 m m glucose for 1 and 8 h, respectively. Phagocytosis of fluorescein-labeled E. coli particles is presented as mean values ( n = 6) ± S.E. of two independent experiments. Phagocytosis of WT cells was arbitrarily set to 100%. *, p

    Article Snippet: Macrophages were cultured in DMEM, 10% LPDS (Invitrogen) supplemented with 1% penicillin, 1% streptomycin for 2 h. Thereafter, cells were washed twice with PBS, and if not stated otherwise, the adherent macrophages were cultured in high glucose (25 mm ) DMEM containing 4 mm glutamine, 1 mm pyruvate, and 10% LPDS.

    Techniques: Mouse Assay, Labeling

    ATP concentrations and ADP/ATP ratios in Atgl −/− and WT macrophages. A , ATP levels in Atgl −/− and WT MPM were measured after cultivating the cells for 24 h in DMEM, 10% LPDS containing 0, 6, and 25 m m glucose. Oligomycin (0.5 μ m ) was used as an ATP-depletion control. Data are presented as mean values ( n = 4) ± S.E. ***, p ≤ 0.001. B , ADP/ATP ratios in Atgl −/− and WT MPM were determined using the EnzyLight TM ADP/ATP ratio assay kit. Data are expressed as mean values ( n = 4) ± S.E. *, p

    Journal: The Journal of Biological Chemistry

    Article Title: Efficient Phagocytosis Requires Triacylglycerol Hydrolysis by Adipose Triglyceride Lipase *

    doi: 10.1074/jbc.M110.107854

    Figure Lengend Snippet: ATP concentrations and ADP/ATP ratios in Atgl −/− and WT macrophages. A , ATP levels in Atgl −/− and WT MPM were measured after cultivating the cells for 24 h in DMEM, 10% LPDS containing 0, 6, and 25 m m glucose. Oligomycin (0.5 μ m ) was used as an ATP-depletion control. Data are presented as mean values ( n = 4) ± S.E. ***, p ≤ 0.001. B , ADP/ATP ratios in Atgl −/− and WT MPM were determined using the EnzyLight TM ADP/ATP ratio assay kit. Data are expressed as mean values ( n = 4) ± S.E. *, p

    Article Snippet: Macrophages were cultured in DMEM, 10% LPDS (Invitrogen) supplemented with 1% penicillin, 1% streptomycin for 2 h. Thereafter, cells were washed twice with PBS, and if not stated otherwise, the adherent macrophages were cultured in high glucose (25 mm ) DMEM containing 4 mm glutamine, 1 mm pyruvate, and 10% LPDS.

    Techniques:

    METTL3 is degraded by caspase activity during lytic replication. (A) TREx-BCBL1-RTA cells were reactivated with 1 µg/mL dox in the presense of absence of 20 ng/mL 12-O-Tetradecanoylphorbol-13-acetate (TPA) and (B) iSLK.219 cells reactivated with 1 µg/mL dox in the presence of absence of sodium butyrate (NaB). METTL3 abundance was assessed by western blot. (C) Uninfected iSLK cells treated with 1 µg/mL dox to stimulate RTA expression were concurrently treated 10 µg/mL MG132 for 24 h and probed as indicated by western blot. (D) TREx-BCBL1-RTA cells treated with 1 µg/mL dox and 10 µM of IDN-6556 to inhibit caspases. Lysates were probed as indicated by western blot.

    Journal: bioRxiv

    Article Title: KSHV lytic mRNA is efficiently translated in the absence of eIF4F

    doi: 10.1101/356162

    Figure Lengend Snippet: METTL3 is degraded by caspase activity during lytic replication. (A) TREx-BCBL1-RTA cells were reactivated with 1 µg/mL dox in the presense of absence of 20 ng/mL 12-O-Tetradecanoylphorbol-13-acetate (TPA) and (B) iSLK.219 cells reactivated with 1 µg/mL dox in the presence of absence of sodium butyrate (NaB). METTL3 abundance was assessed by western blot. (C) Uninfected iSLK cells treated with 1 µg/mL dox to stimulate RTA expression were concurrently treated 10 µg/mL MG132 for 24 h and probed as indicated by western blot. (D) TREx-BCBL1-RTA cells treated with 1 µg/mL dox and 10 µM of IDN-6556 to inhibit caspases. Lysates were probed as indicated by western blot.

    Article Snippet: All cells were maintained in 100 I/U of both penicillin and streptomycin. iSLK.219 cells were cultured with 10 mM puromycin (ThermoFisher Scientific) to maintain episome copy number of rKSHV.219.

    Techniques: Activity Assay, Western Blot, Expressing

    KSHV lytic replication reduces protein synthesis. (A) Western blots of whole cell lysates for polysome profiles. (B) Polysome profiles of uninfected, latent, or 48 hpi iSLK.219 treated with either Torin or DMSO for 2 h prior to harvest. Cells were treated with 100 µg/mL cycloheximide (CHX) for 5 minutes prior to harvest to prevent elongation. Cells were lysed in the presence of CHX and loaded on a 7-47% linear sucrose gradient. After separation by ultracentrifugation, the abundance of RNA (A 260 nm) in the gradient was continually measure as fractions were collected. RNA from the 48 hpi polysome fractions was isolated for sequencing.

    Journal: bioRxiv

    Article Title: KSHV lytic mRNA is efficiently translated in the absence of eIF4F

    doi: 10.1101/356162

    Figure Lengend Snippet: KSHV lytic replication reduces protein synthesis. (A) Western blots of whole cell lysates for polysome profiles. (B) Polysome profiles of uninfected, latent, or 48 hpi iSLK.219 treated with either Torin or DMSO for 2 h prior to harvest. Cells were treated with 100 µg/mL cycloheximide (CHX) for 5 minutes prior to harvest to prevent elongation. Cells were lysed in the presence of CHX and loaded on a 7-47% linear sucrose gradient. After separation by ultracentrifugation, the abundance of RNA (A 260 nm) in the gradient was continually measure as fractions were collected. RNA from the 48 hpi polysome fractions was isolated for sequencing.

    Article Snippet: All cells were maintained in 100 I/U of both penicillin and streptomycin. iSLK.219 cells were cultured with 10 mM puromycin (ThermoFisher Scientific) to maintain episome copy number of rKSHV.219.

    Techniques: Western Blot, Isolation, Sequencing

    2-AI increased penicillin V and vancomycin binding to M . tuberculosis . A and B) Representative histogram overlays from flow cytometry analysis (right panels) of BOCILLIN ® or BODIPY ® FL vancomycin stained M . tuberculosis after treatment with 125 μM 2B8 or DMSO control. Fold-change staining (left panels) was determined by dividing percent of positively-stained treated bacteria by percent of positively-stained DMSO control bacteria. For BOCILLIN ® , a time-dependent increase in staining was observed with 2B8, but not with RA11, SDS or MCA treatment. 2B8 treatment also increased staining with BODIPY ® FL vancomycin to a significantly higher extent in a time-dependent manner compared to RA11 or SDS treatment. MCA treatment resulted in a time-dependent increase of BODIPY ® FL vancomycin staining as expected. C) Fluorescent images (×1000) of M . tuberculosis stained with BODIPY ® FL vancomycin after treatment or not with 125 μM 2B8 for 120 min. Treatment with 2B8 (right panel) resulted in increased number of bacteria with higher fluorescence intensity than DMSO control. *p

    Journal: PLoS ONE

    Article Title: 2-aminoimidazoles potentiate ß-lactam antimicrobial activity against Mycobacterium tuberculosis by reducing ß-lactamase secretion and increasing cell envelope permeability

    doi: 10.1371/journal.pone.0180925

    Figure Lengend Snippet: 2-AI increased penicillin V and vancomycin binding to M . tuberculosis . A and B) Representative histogram overlays from flow cytometry analysis (right panels) of BOCILLIN ® or BODIPY ® FL vancomycin stained M . tuberculosis after treatment with 125 μM 2B8 or DMSO control. Fold-change staining (left panels) was determined by dividing percent of positively-stained treated bacteria by percent of positively-stained DMSO control bacteria. For BOCILLIN ® , a time-dependent increase in staining was observed with 2B8, but not with RA11, SDS or MCA treatment. 2B8 treatment also increased staining with BODIPY ® FL vancomycin to a significantly higher extent in a time-dependent manner compared to RA11 or SDS treatment. MCA treatment resulted in a time-dependent increase of BODIPY ® FL vancomycin staining as expected. C) Fluorescent images (×1000) of M . tuberculosis stained with BODIPY ® FL vancomycin after treatment or not with 125 μM 2B8 for 120 min. Treatment with 2B8 (right panel) resulted in increased number of bacteria with higher fluorescence intensity than DMSO control. *p

    Article Snippet: After treating for 30 min, 120 min, or 24 h at 37°C while shaking, cultures were aliquoted into 5 mL polystyrene tubes and stained with 1 mg/L BODIPY® FL vancomycin, 10 mg/L BODIPY® -tagged penicillin V (BOCILLIN®) ) or 15 μM propidium iodide (PI) (Life Technologies, Carlsbad, CA, USA), for 30 min at 37°C in the dark.

    Techniques: Binding Assay, Flow Cytometry, Cytometry, Staining, Fluorescence

    Human T cell proliferation was affected by edelfosine. (A) Reduced PBMC proliferation upon addition of edelfosine on cell seeding was independent of the addition of PHA. Notably, PHA-activated cells appeared to be susceptible to edelfosine at 10-fold lower concentrations. (B) The inhibitory effect of edelfosine was observed if the drug was added to already activated, proliferating T cells, i.e. two days after cell seeding and PHA addition. Here, a significant reduction of proliferation in unstimulated cells was only detectable with 33.3 µg/ml edelfosine. (C) Preincubation of PBMCs with at least 3.3 µg/ml edelfosine interfered with the cells' capacity to proliferate upon PHA stimulation. No effect was detected in preconditioned, but unstimulated cells (experiments A, B, C: sample size n = 3 donors, each approach was seeded in triplicates and means for each donor are represented by symbols •, ▪, ▴). (D) 1 µg/ml edelfosine or higher concentrations profoundly diminished proliferation in MBP (83–99) -specific TCLs. One representative TCL of two is shown. Cells were incubated in quadruplicates. • stimulated, ▪ unstimulated (E) PBMCs of one donor were cultured without addition of a stimulus. Proliferation was detectable after seven days. The presence of anti-HLA-DR- and anti-MHC class I-blocking antibodies or 3.3 µg/ml edelfosine inhibited cellular proliferation (• untreated, ▪ blocking antibodies added, ▴ 3.3 µg/ml edelfosine-treated). Bars represent mean values ± SEM, *P

    Journal: PLoS ONE

    Article Title: The Orally Available, Synthetic Ether Lipid Edelfosine Inhibits T Cell Proliferation and Induces a Type I Interferon Response

    doi: 10.1371/journal.pone.0091970

    Figure Lengend Snippet: Human T cell proliferation was affected by edelfosine. (A) Reduced PBMC proliferation upon addition of edelfosine on cell seeding was independent of the addition of PHA. Notably, PHA-activated cells appeared to be susceptible to edelfosine at 10-fold lower concentrations. (B) The inhibitory effect of edelfosine was observed if the drug was added to already activated, proliferating T cells, i.e. two days after cell seeding and PHA addition. Here, a significant reduction of proliferation in unstimulated cells was only detectable with 33.3 µg/ml edelfosine. (C) Preincubation of PBMCs with at least 3.3 µg/ml edelfosine interfered with the cells' capacity to proliferate upon PHA stimulation. No effect was detected in preconditioned, but unstimulated cells (experiments A, B, C: sample size n = 3 donors, each approach was seeded in triplicates and means for each donor are represented by symbols •, ▪, ▴). (D) 1 µg/ml edelfosine or higher concentrations profoundly diminished proliferation in MBP (83–99) -specific TCLs. One representative TCL of two is shown. Cells were incubated in quadruplicates. • stimulated, ▪ unstimulated (E) PBMCs of one donor were cultured without addition of a stimulus. Proliferation was detectable after seven days. The presence of anti-HLA-DR- and anti-MHC class I-blocking antibodies or 3.3 µg/ml edelfosine inhibited cellular proliferation (• untreated, ▪ blocking antibodies added, ▴ 3.3 µg/ml edelfosine-treated). Bars represent mean values ± SEM, *P

    Article Snippet: In brief, 2×105 PBMCs per well were seeded in 96-well plates (Greiner Bio-One) in complete TCL medium (1% penicillin/streptomycin (Gibco), 0.05 g/l gentamycin (Lonza), 2 mM L-glutamine (Gibco), 5% human serum (PAA) in RPMI1640 medium, GlutaMAX (Gibco)) supplemented with 10 µg/ml MBP(83–99) peptide (peptides & elephants).

    Techniques: Incubation, Cell Culture, Blocking Assay