medium dmem Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    ATCC dulbeccos s modified eagle s medium
    Dulbeccos S Modified Eagle S Medium, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dulbeccos s modified eagle s medium/product/ATCC
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    dulbeccos s modified eagle s medium - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher dulbeccos s modified eagle s medium dmem
    Dulbeccos S Modified Eagle S Medium Dmem, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dulbeccos s modified eagle s medium dmem/product/Thermo Fisher
    Average 99 stars, based on 34 article reviews
    Price from $9.99 to $1999.99
    dulbeccos s modified eagle s medium dmem - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    90
    GE Healthcare dulbeccos s modified eagle s medium dmem
    Dulbeccos S Modified Eagle S Medium Dmem, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 90/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dulbeccos s modified eagle s medium dmem/product/GE Healthcare
    Average 90 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    dulbeccos s modified eagle s medium dmem - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    99
    Thermo Fisher dulbeccos s modified eagle s medium
    Dulbeccos S Modified Eagle S Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dulbeccos s modified eagle s medium/product/Thermo Fisher
    Average 99 stars, based on 51 article reviews
    Price from $9.99 to $1999.99
    dulbeccos s modified eagle s medium - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    92
    Lonza complete dulbeccos s modified eagle s medium dmem
    Complete Dulbeccos S Modified Eagle S Medium Dmem, supplied by Lonza, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/complete dulbeccos s modified eagle s medium dmem/product/Lonza
    Average 92 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    complete dulbeccos s modified eagle s medium dmem - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    99
    Millipore glucose dulbeccos s modified eagle s medium
    Glucose Dulbeccos S Modified Eagle S Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glucose dulbeccos s modified eagle s medium/product/Millipore
    Average 99 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    glucose dulbeccos s modified eagle s medium - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    88
    Mediatech dulbeccos s modified eagle s medium
    Dulbeccos S Modified Eagle S Medium, supplied by Mediatech, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dulbeccos s modified eagle s medium/product/Mediatech
    Average 88 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    dulbeccos s modified eagle s medium - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    99
    Thermo Fisher dmem medium dmem
    Inhibition of EV71 infection by E151 at the postattachment stage. (A) E151 inhibited EV71-GFP infection following attachment. Prechilled RD or <t>Vero</t> cells were incubated with ice-cold EV71-GFP at an MOI of 10 and attachment allowed at 4°C for 1 h. The unattached viruses were then washed away from cells using ice-cold PBS. Fresh <t>DMEM-10</t> with or without 100 μM E151 was added, and the cells were transferred into a 37°C incubator. After 11 h, the cells were fixed and stained with Hoechst 33258. The percentage of infected cells (green) at 3 random spots under each condition is presented as the average value ± standard error in the top left corner of a representative image. (B) E151 detached cell-bound RG/EV71-B5-KEP but not RG/EV71-B5-KEA in the immunofluorescence confocal microscopy assay. Attached virions were allowed to internalize into the cells at 37°C for 30 min in the presence or absence of 100 μM E151. Virions (green), cellular endosomes (red), and cellular DNA (cyan) were stained by anti-EV71 antiserum, anti-EEA1 antibody, and Hoechst 33258, respectively, after cell fixation. The numbers of EV71 puncta per cell (average values ± standard errors) in representative green EV71 channel images (×600 magnification) are presented in the top right corners. Enlarged views of the small boxed areas in the corresponding multiple EV71/EEA1/DNA channel images are in the top left corners. (C) Attached EV71 virions were eluted from RD cells by E151 in a concentration-dependent manner. E151-sensitive EV71-B5 or -resistant EV71-B5 res was incubated with RD cells at 4°C for 1 h for attachment, and unattached viruses were washed away with ice-cold PBS. The attached viruses-cells were divided into 3 portions and resuspended in ice-cold PBS with 0, 30, or 100 μM E151. After incubation at 4°C for 15 min with gentle agitation, the cell pellets and supernatants were separated for EV71 antigen detection using Western blotting. (D) EV71 binding to the uncoating factor cyclophilin A (CypA) was prevented by E151. In the GST pulldown assay, glutathione-Sepharose-bound GST or GST-CypA was incubated with purified EV71-B5 or EV71-E5 res in the absence or presence of E151. The precipitated proteins were analyzed by Western blotting with the antibody 1D9 and anti-GST antiserum.
    Dmem Medium Dmem, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3165 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dmem medium dmem/product/Thermo Fisher
    Average 99 stars, based on 3165 article reviews
    Price from $9.99 to $1999.99
    dmem medium dmem - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    Image Search Results


    Inhibition of EV71 infection by E151 at the postattachment stage. (A) E151 inhibited EV71-GFP infection following attachment. Prechilled RD or Vero cells were incubated with ice-cold EV71-GFP at an MOI of 10 and attachment allowed at 4°C for 1 h. The unattached viruses were then washed away from cells using ice-cold PBS. Fresh DMEM-10 with or without 100 μM E151 was added, and the cells were transferred into a 37°C incubator. After 11 h, the cells were fixed and stained with Hoechst 33258. The percentage of infected cells (green) at 3 random spots under each condition is presented as the average value ± standard error in the top left corner of a representative image. (B) E151 detached cell-bound RG/EV71-B5-KEP but not RG/EV71-B5-KEA in the immunofluorescence confocal microscopy assay. Attached virions were allowed to internalize into the cells at 37°C for 30 min in the presence or absence of 100 μM E151. Virions (green), cellular endosomes (red), and cellular DNA (cyan) were stained by anti-EV71 antiserum, anti-EEA1 antibody, and Hoechst 33258, respectively, after cell fixation. The numbers of EV71 puncta per cell (average values ± standard errors) in representative green EV71 channel images (×600 magnification) are presented in the top right corners. Enlarged views of the small boxed areas in the corresponding multiple EV71/EEA1/DNA channel images are in the top left corners. (C) Attached EV71 virions were eluted from RD cells by E151 in a concentration-dependent manner. E151-sensitive EV71-B5 or -resistant EV71-B5 res was incubated with RD cells at 4°C for 1 h for attachment, and unattached viruses were washed away with ice-cold PBS. The attached viruses-cells were divided into 3 portions and resuspended in ice-cold PBS with 0, 30, or 100 μM E151. After incubation at 4°C for 15 min with gentle agitation, the cell pellets and supernatants were separated for EV71 antigen detection using Western blotting. (D) EV71 binding to the uncoating factor cyclophilin A (CypA) was prevented by E151. In the GST pulldown assay, glutathione-Sepharose-bound GST or GST-CypA was incubated with purified EV71-B5 or EV71-E5 res in the absence or presence of E151. The precipitated proteins were analyzed by Western blotting with the antibody 1D9 and anti-GST antiserum.

    Journal: Journal of Virology

    Article Title: In Vitro and In Vivo Inhibition of the Infectivity of Human Enterovirus 71 by a Sulfonated Food Azo Dye, Brilliant Black BN

    doi: 10.1128/JVI.00061-19

    Figure Lengend Snippet: Inhibition of EV71 infection by E151 at the postattachment stage. (A) E151 inhibited EV71-GFP infection following attachment. Prechilled RD or Vero cells were incubated with ice-cold EV71-GFP at an MOI of 10 and attachment allowed at 4°C for 1 h. The unattached viruses were then washed away from cells using ice-cold PBS. Fresh DMEM-10 with or without 100 μM E151 was added, and the cells were transferred into a 37°C incubator. After 11 h, the cells were fixed and stained with Hoechst 33258. The percentage of infected cells (green) at 3 random spots under each condition is presented as the average value ± standard error in the top left corner of a representative image. (B) E151 detached cell-bound RG/EV71-B5-KEP but not RG/EV71-B5-KEA in the immunofluorescence confocal microscopy assay. Attached virions were allowed to internalize into the cells at 37°C for 30 min in the presence or absence of 100 μM E151. Virions (green), cellular endosomes (red), and cellular DNA (cyan) were stained by anti-EV71 antiserum, anti-EEA1 antibody, and Hoechst 33258, respectively, after cell fixation. The numbers of EV71 puncta per cell (average values ± standard errors) in representative green EV71 channel images (×600 magnification) are presented in the top right corners. Enlarged views of the small boxed areas in the corresponding multiple EV71/EEA1/DNA channel images are in the top left corners. (C) Attached EV71 virions were eluted from RD cells by E151 in a concentration-dependent manner. E151-sensitive EV71-B5 or -resistant EV71-B5 res was incubated with RD cells at 4°C for 1 h for attachment, and unattached viruses were washed away with ice-cold PBS. The attached viruses-cells were divided into 3 portions and resuspended in ice-cold PBS with 0, 30, or 100 μM E151. After incubation at 4°C for 15 min with gentle agitation, the cell pellets and supernatants were separated for EV71 antigen detection using Western blotting. (D) EV71 binding to the uncoating factor cyclophilin A (CypA) was prevented by E151. In the GST pulldown assay, glutathione-Sepharose-bound GST or GST-CypA was incubated with purified EV71-B5 or EV71-E5 res in the absence or presence of E151. The precipitated proteins were analyzed by Western blotting with the antibody 1D9 and anti-GST antiserum.

    Article Snippet: Human rhabdomyosarcoma (RD; ATCC number CCL-136) and monkey Vero (ATCC number CCL-81) cell lines were maintained in Dulbecco’s modified Eagles’ medium (DMEM) (Life Technologies) supplemented with 10% fetal bovine serum (FBS) (Biowest) and 1× antibiotic-antimycotic (Life Technologies) at 37°C in a 5% CO2 incubator.

    Techniques: Inhibition, Infection, Incubation, Staining, Immunofluorescence, Confocal Microscopy, Concentration Assay, Western Blot, Binding Assay, GST Pulldown Assay, Purification

    Analysis of c-Fos and c-Jun protein expression in cervical cancer cells. Total cellular proteins were obtained from 1 × 10 5 HaCaT (lane 1), C-33A (lane 2), SiHa (lane 3) and HeLa cells (lane 4) per well in a six-well plate containing DMEM at 37 °C with 10% FBS in 5% CO 2 . The proteins were separated in 12% SDS-PAGE and were transferred to nitrocellulose membranes, which were incubated with each antibody. a The amount of similar proteins analyzed in the immunoblots. The anti-beta-actin antibody was included as control. b The immunoblot bands digitalized and analyzed by densitometer. The data were analyzed by c-Fos/c-Jun/beta-actin fold change in relative expression units (mean ± SE), not statistically significant (ns) and P values

    Journal: Cancer Cell International

    Article Title: Transregulation of microRNA miR-21 promoter by AP-1 transcription factor in cervical cancer cells

    doi: 10.1186/s12935-019-0931-x

    Figure Lengend Snippet: Analysis of c-Fos and c-Jun protein expression in cervical cancer cells. Total cellular proteins were obtained from 1 × 10 5 HaCaT (lane 1), C-33A (lane 2), SiHa (lane 3) and HeLa cells (lane 4) per well in a six-well plate containing DMEM at 37 °C with 10% FBS in 5% CO 2 . The proteins were separated in 12% SDS-PAGE and were transferred to nitrocellulose membranes, which were incubated with each antibody. a The amount of similar proteins analyzed in the immunoblots. The anti-beta-actin antibody was included as control. b The immunoblot bands digitalized and analyzed by densitometer. The data were analyzed by c-Fos/c-Jun/beta-actin fold change in relative expression units (mean ± SE), not statistically significant (ns) and P values

    Article Snippet: The cell lines were cultured in DMEM medium (Invitrogen, Carlsbad, CA) with 10% FBS, 100 U/ml penicillin/100 μg/ml streptomycin, 2 mM l -glutamine, 250 ng/ml fungizone, and maintained at 37 °C in 5% CO2.

    Techniques: Expressing, SDS Page, Incubation, Western Blot

    MiR-21 gene expression analysis after constitutive activation of AP-1. Total RNA and cDNA synthesis were obtained from 1 × 10 5 SiHa and HaCaT cells per well in a six-well plate containing DMEM at 37 °C with 10% FBS in 5% CO 2 after 60 min of non-treatment (NT) or treatment with 10 ng/ml PMA and 50 μM SR11302. The miR-21 gene expression was analyzed by real-time qRT-PCR and relative expression of miR-21 was calculated using the 2 −∆∆Ct method. The RNU44 gene expression was used as control. The data were analyzed by mRNA miR-21/mRNA RNU44 ratio in relative expression units. The values are presented as mean ± SD and P values

    Journal: Cancer Cell International

    Article Title: Transregulation of microRNA miR-21 promoter by AP-1 transcription factor in cervical cancer cells

    doi: 10.1186/s12935-019-0931-x

    Figure Lengend Snippet: MiR-21 gene expression analysis after constitutive activation of AP-1. Total RNA and cDNA synthesis were obtained from 1 × 10 5 SiHa and HaCaT cells per well in a six-well plate containing DMEM at 37 °C with 10% FBS in 5% CO 2 after 60 min of non-treatment (NT) or treatment with 10 ng/ml PMA and 50 μM SR11302. The miR-21 gene expression was analyzed by real-time qRT-PCR and relative expression of miR-21 was calculated using the 2 −∆∆Ct method. The RNU44 gene expression was used as control. The data were analyzed by mRNA miR-21/mRNA RNU44 ratio in relative expression units. The values are presented as mean ± SD and P values

    Article Snippet: The cell lines were cultured in DMEM medium (Invitrogen, Carlsbad, CA) with 10% FBS, 100 U/ml penicillin/100 μg/ml streptomycin, 2 mM l -glutamine, 250 ng/ml fungizone, and maintained at 37 °C in 5% CO2.

    Techniques: Expressing, Activation Assay, Quantitative RT-PCR

    Analysis of c-Fos and c-Jun protein expression after constitutive activation of AP-1. Total cellular proteins were obtained from 1 × 10 5 SiHa ( a ) and HaCaT cells ( b ) per well in a six-well plate containing DMEM at 37 °C with 10% FBS in 5% CO 2 after 0, 24, 48 and 72 h treated with 10 ng/ml PMA (lanes 1–4) or 50 μM SR11302 (lanes 5–8). The proteins were separated in 12% SDS-PAGE and were transferred to nitrocellulose membranes, which were incubated with each antibody. Similar amount of proteins were analyzed in the immunoblots. The anti-beta-actin antibody was included as control. The immunoblot bands were digitalized and analyzed by densitometer. The data were analyzed by c-Fos/c-Jun/beta-actin fold change in relative expression units (mean ± SE), not statistically significant (ns) and P values

    Journal: Cancer Cell International

    Article Title: Transregulation of microRNA miR-21 promoter by AP-1 transcription factor in cervical cancer cells

    doi: 10.1186/s12935-019-0931-x

    Figure Lengend Snippet: Analysis of c-Fos and c-Jun protein expression after constitutive activation of AP-1. Total cellular proteins were obtained from 1 × 10 5 SiHa ( a ) and HaCaT cells ( b ) per well in a six-well plate containing DMEM at 37 °C with 10% FBS in 5% CO 2 after 0, 24, 48 and 72 h treated with 10 ng/ml PMA (lanes 1–4) or 50 μM SR11302 (lanes 5–8). The proteins were separated in 12% SDS-PAGE and were transferred to nitrocellulose membranes, which were incubated with each antibody. Similar amount of proteins were analyzed in the immunoblots. The anti-beta-actin antibody was included as control. The immunoblot bands were digitalized and analyzed by densitometer. The data were analyzed by c-Fos/c-Jun/beta-actin fold change in relative expression units (mean ± SE), not statistically significant (ns) and P values

    Article Snippet: The cell lines were cultured in DMEM medium (Invitrogen, Carlsbad, CA) with 10% FBS, 100 U/ml penicillin/100 μg/ml streptomycin, 2 mM l -glutamine, 250 ng/ml fungizone, and maintained at 37 °C in 5% CO2.

    Techniques: Expressing, Activation Assay, SDS Page, Incubation, Western Blot

    Phenotypic characterization of M. bovis vaccine strain P150. EBL cell survival (% OD590 nm) after 72 h of co-incubation with HB0801 (P1) and P150 (P150) in DMEM-based medium supplemented with 10 μg/ml calf thymus DNA (A) . Mycoplasma titers (log CFU/ml) reached by P1 and P150 after 48 h incubation in axenic DMEM-based medium without pyruvate and supplemented with 10 μg/ml calf thymus DNA and H 2 O 2 production as determined by using MQuant ™ Peroxid-test strips (B) . Data are the means of at least three independent assays. Standard deviations are indicated by error bars.

    Journal: Frontiers in Microbiology

    Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity

    doi: 10.3389/fmicb.2019.02753

    Figure Lengend Snippet: Phenotypic characterization of M. bovis vaccine strain P150. EBL cell survival (% OD590 nm) after 72 h of co-incubation with HB0801 (P1) and P150 (P150) in DMEM-based medium supplemented with 10 μg/ml calf thymus DNA (A) . Mycoplasma titers (log CFU/ml) reached by P1 and P150 after 48 h incubation in axenic DMEM-based medium without pyruvate and supplemented with 10 μg/ml calf thymus DNA and H 2 O 2 production as determined by using MQuant ™ Peroxid-test strips (B) . Data are the means of at least three independent assays. Standard deviations are indicated by error bars.

    Article Snippet: EBL cells were seeded at a density of 5 × 103 cells/cm2 in DMEM-based medium supplemented with 10 μg/ml calf thymus DNA (Invitrogen) and inoculated with individual mutants using the 96-pin replicator (Boekel Scientific).

    Techniques: Incubation

    The growth-promoting effect of eDNA on M. bovis . Comparative growth of M. bovis under axenic and cell culture conditions (A) . RM16 proliferation was monitored in SP4 medium (SP4), cell culture medium (DMEM), and cell culture medium supplemented with 10 μg/ml calf thymus DNA (DMEMD). Mycoplasma titers (log CFU/ml) were determined by CFU titrations. The cytopathic effect induced by M. bovis upon co-incubation with host cells (B) . EBL cells (10 4 cells) were inoculated with RM16 at an MOI of 2 (RM16) or mock-infected (Mock). After 72 h of co-incubation, monolayers were stained with crystal violet and survival cells were estimated by measuring the optical density at 590 nm (OD 590). When indicated, DMEM-based medium was supplemented with 10 μg/ml calf thymus DNA (eDNA). Calf thymus DNA was subjected to the following enzymatic treatments: RNase A (RNase), Proteinase K (ProtK) and DNase I (DNase) digestion (see section “Materials and Methods”). The asterisk indicates that polynucleotides were removed from DNase I digestion products (DNase*). Infected and mock-infected samples were treated identically. Data are the means of at least three independent assays. Standard deviations are indicated by error bars. p -values were determined by using two-sided independent sample t tests and comparing OD590 values of RM16-infected samples to those of mock-infected samples ( *** p

    Journal: Frontiers in Microbiology

    Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity

    doi: 10.3389/fmicb.2019.02753

    Figure Lengend Snippet: The growth-promoting effect of eDNA on M. bovis . Comparative growth of M. bovis under axenic and cell culture conditions (A) . RM16 proliferation was monitored in SP4 medium (SP4), cell culture medium (DMEM), and cell culture medium supplemented with 10 μg/ml calf thymus DNA (DMEMD). Mycoplasma titers (log CFU/ml) were determined by CFU titrations. The cytopathic effect induced by M. bovis upon co-incubation with host cells (B) . EBL cells (10 4 cells) were inoculated with RM16 at an MOI of 2 (RM16) or mock-infected (Mock). After 72 h of co-incubation, monolayers were stained with crystal violet and survival cells were estimated by measuring the optical density at 590 nm (OD 590). When indicated, DMEM-based medium was supplemented with 10 μg/ml calf thymus DNA (eDNA). Calf thymus DNA was subjected to the following enzymatic treatments: RNase A (RNase), Proteinase K (ProtK) and DNase I (DNase) digestion (see section “Materials and Methods”). The asterisk indicates that polynucleotides were removed from DNase I digestion products (DNase*). Infected and mock-infected samples were treated identically. Data are the means of at least three independent assays. Standard deviations are indicated by error bars. p -values were determined by using two-sided independent sample t tests and comparing OD590 values of RM16-infected samples to those of mock-infected samples ( *** p

    Article Snippet: EBL cells were seeded at a density of 5 × 103 cells/cm2 in DMEM-based medium supplemented with 10 μg/ml calf thymus DNA (Invitrogen) and inoculated with individual mutants using the 96-pin replicator (Boekel Scientific).

    Techniques: Cell Culture, Incubation, Infection, Staining

    Phenotypic characterization of M. bovis knockout mutants (A) and M. bovis type and field strains (B) . M. bovis mutants and strains are described in Tables 1 , 2 . Mycoplasmas were co-incubated with EBL cells in DMEM-based medium supplemented with 10 μg/ml calf thymus DNA (black bars) or DMEM-based medium without DNA (white bars). EBL cells (10 4 cells) were inoculated at an MOI of 2. After 72 h of co-incubation, samples were used either for CFU titrations (log CFU/ml), analysis of EBL cell survival (% OD590 nm) or hydrogen peroxide production (H 2 O 2 ). Infected and mock-infected (Mock) samples were treated identically. Data are the means of at least three independent assays. Standard deviations are indicated by error bars. p -values were determined by using two-sided independent sample t tests and comparing (1) mycoplasma titers (log CFU/ml) reached in the presence of DNA to those reached without supplementation, and (2) EBL cell survival (% OD590 nm) or (3) H2O2 production (H2O2) following co-incubation with M. bovis mutants or strains to values obtained with RM16 (ns, p > 0.05; ** p

    Journal: Frontiers in Microbiology

    Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity

    doi: 10.3389/fmicb.2019.02753

    Figure Lengend Snippet: Phenotypic characterization of M. bovis knockout mutants (A) and M. bovis type and field strains (B) . M. bovis mutants and strains are described in Tables 1 , 2 . Mycoplasmas were co-incubated with EBL cells in DMEM-based medium supplemented with 10 μg/ml calf thymus DNA (black bars) or DMEM-based medium without DNA (white bars). EBL cells (10 4 cells) were inoculated at an MOI of 2. After 72 h of co-incubation, samples were used either for CFU titrations (log CFU/ml), analysis of EBL cell survival (% OD590 nm) or hydrogen peroxide production (H 2 O 2 ). Infected and mock-infected (Mock) samples were treated identically. Data are the means of at least three independent assays. Standard deviations are indicated by error bars. p -values were determined by using two-sided independent sample t tests and comparing (1) mycoplasma titers (log CFU/ml) reached in the presence of DNA to those reached without supplementation, and (2) EBL cell survival (% OD590 nm) or (3) H2O2 production (H2O2) following co-incubation with M. bovis mutants or strains to values obtained with RM16 (ns, p > 0.05; ** p

    Article Snippet: EBL cells were seeded at a density of 5 × 103 cells/cm2 in DMEM-based medium supplemented with 10 μg/ml calf thymus DNA (Invitrogen) and inoculated with individual mutants using the 96-pin replicator (Boekel Scientific).

    Techniques: Knock-Out, Incubation, Infection

    Saliva does not inactivate HSV-1 amplicon particles (A) A HSV-1 amplicon vector encoding luciferase was suspended in DMEM medium without FBS and incubated with an equal volume of whole unstimulated human saliva at 37°C for 30 minutes. Following incubation, complete DMEM (containing 10% FBS plus antibiotics) was added and the mixture was then added to HEK 293A cells. Twenty-four hours later the 293A cells were washed with PBS, lysed with passive lysis buffer (Promega), centrifuged at 13,000 RPM for 10 minutes to remove cell debris, and then the lysate was directly used to quantitate luciferase expression. Luciferase expression levels in cells exposed to control virus stocks (not incubated with saliva) was assigned a level of 100%, and luciferase levels in cells exposed to saliva-treated virus stocks was calculated as a percentage of this baseline. The results shown represent mean normalized luciferase expression data (% of baseline relative light units; RLU) from virus samples that were incubated with each of four different human saliva samples; error bars denote the standard error of the mean. (B) A HSV-1 amplicon vector encoding GFP was used to conduct an experiment analogous to that described for panel A. In this case, the vector was preincubated with pooled, pilocarpine-stimulated whole mouse saliva (collected from three female BALB/c mice). 24 hours following addition of control or saliva-treated vector to 293A cell cultures, the cells were collected and analyzed by flow cytometry. The percentage of GFP positive cells in cultures exposed to control virus stocks (not incubated with saliva) was assigned a level of 100%, and the transduction efficiency in cells exposed to saliva-treated virus stocks was calculated as a percentage of this baseline. The results shown represent the mean normalized transduction efficiency data calculated from four replicates; error bars denote the standard error of the mean.

    Journal: Vaccine

    Article Title: Robust Antigen-Specific Humoral Immune Responses to Sublingually Delivered Adenoviral Vectors Encoding HIV-1 Env: Association with Mucoadhesion and Efficient Penetration of the Sublingual Barrier

    doi: 10.1016/j.vaccine.2011.07.008

    Figure Lengend Snippet: Saliva does not inactivate HSV-1 amplicon particles (A) A HSV-1 amplicon vector encoding luciferase was suspended in DMEM medium without FBS and incubated with an equal volume of whole unstimulated human saliva at 37°C for 30 minutes. Following incubation, complete DMEM (containing 10% FBS plus antibiotics) was added and the mixture was then added to HEK 293A cells. Twenty-four hours later the 293A cells were washed with PBS, lysed with passive lysis buffer (Promega), centrifuged at 13,000 RPM for 10 minutes to remove cell debris, and then the lysate was directly used to quantitate luciferase expression. Luciferase expression levels in cells exposed to control virus stocks (not incubated with saliva) was assigned a level of 100%, and luciferase levels in cells exposed to saliva-treated virus stocks was calculated as a percentage of this baseline. The results shown represent mean normalized luciferase expression data (% of baseline relative light units; RLU) from virus samples that were incubated with each of four different human saliva samples; error bars denote the standard error of the mean. (B) A HSV-1 amplicon vector encoding GFP was used to conduct an experiment analogous to that described for panel A. In this case, the vector was preincubated with pooled, pilocarpine-stimulated whole mouse saliva (collected from three female BALB/c mice). 24 hours following addition of control or saliva-treated vector to 293A cell cultures, the cells were collected and analyzed by flow cytometry. The percentage of GFP positive cells in cultures exposed to control virus stocks (not incubated with saliva) was assigned a level of 100%, and the transduction efficiency in cells exposed to saliva-treated virus stocks was calculated as a percentage of this baseline. The results shown represent the mean normalized transduction efficiency data calculated from four replicates; error bars denote the standard error of the mean.

    Article Snippet: The virus/saliva mixture was incubated at 37°C for 30 minutes, after which complete DMEM medium (DMEM [Gibco-BRL] plus 10% fetal bovine serum [FBS] and 1% penicillin, streptomycin and glutamine [PSG]) was added and the mixture was transferred to cultures of HEK 293A cells.

    Techniques: Amplification, Plasmid Preparation, Luciferase, Incubation, Lysis, Expressing, Mouse Assay, Flow Cytometry, Cytometry, Transduction