nmuli  (ATCC)


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    Structured Review

    ATCC nmuli
    F. novicida invades and replicates <t>within</t> <t>BNL</t> CL.2 and <t>NMuLi</t> hepatocytes. (A) Immunofluorescent images of F. novicida infections in cultured hepatocytes. Hepatocytes were fixed following 24 h F. novicida infections with both BNL CL.2 and NMuLi cells. Samples were labelled with anti- F. novicida antibodies (green), fluorescent phalloidin to indicate the cell boundaries (red) and DAPI (blue). Arrowheads indicate some of the bacteria within the infected cells. Scale bar = 10m. (B) Quantification of the proportion of infected hepatocytes as assessed by microscopic examination. NMuLi (n = 17) and BNL CL.2 (n = 16). (C) Titre of intracellular F. novicida at various timepoints following infection of BNL CL.2 and NMuLi cell infections by invasion assay (n = 3).
    Nmuli, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nmuli/product/ATCC
    Average 94 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    nmuli - by Bioz Stars, 2022-09
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    Images

    1) Product Images from "Francisellatularensis Uses Cholesterol and Clathrin-Based Endocytic Mechanisms to Invade Hepatocytes"

    Article Title: Francisellatularensis Uses Cholesterol and Clathrin-Based Endocytic Mechanisms to Invade Hepatocytes

    Journal: Scientific Reports

    doi: 10.1038/srep00192

    F. novicida invades and replicates within BNL CL.2 and NMuLi hepatocytes. (A) Immunofluorescent images of F. novicida infections in cultured hepatocytes. Hepatocytes were fixed following 24 h F. novicida infections with both BNL CL.2 and NMuLi cells. Samples were labelled with anti- F. novicida antibodies (green), fluorescent phalloidin to indicate the cell boundaries (red) and DAPI (blue). Arrowheads indicate some of the bacteria within the infected cells. Scale bar = 10m. (B) Quantification of the proportion of infected hepatocytes as assessed by microscopic examination. NMuLi (n = 17) and BNL CL.2 (n = 16). (C) Titre of intracellular F. novicida at various timepoints following infection of BNL CL.2 and NMuLi cell infections by invasion assay (n = 3).
    Figure Legend Snippet: F. novicida invades and replicates within BNL CL.2 and NMuLi hepatocytes. (A) Immunofluorescent images of F. novicida infections in cultured hepatocytes. Hepatocytes were fixed following 24 h F. novicida infections with both BNL CL.2 and NMuLi cells. Samples were labelled with anti- F. novicida antibodies (green), fluorescent phalloidin to indicate the cell boundaries (red) and DAPI (blue). Arrowheads indicate some of the bacteria within the infected cells. Scale bar = 10m. (B) Quantification of the proportion of infected hepatocytes as assessed by microscopic examination. NMuLi (n = 17) and BNL CL.2 (n = 16). (C) Titre of intracellular F. novicida at various timepoints following infection of BNL CL.2 and NMuLi cell infections by invasion assay (n = 3).

    Techniques Used: Cell Culture, Infection, Invasion Assay

    F. novicida entry does not utilize caveolin-dependent or macropinocytosis pathways. (A) BNL CL.2 and NMuLi cells were treated with the caveolin-inhibitor filipin and infected with F. novicida for 22 h followed by 2 h gentamicin treatment for invasion assays. (B) Immunolocalization of caveolin-1 (green) and DAPI (blue) of uninfected BNL CL.2 cells or following 8 h, 16 h, and 24 h infections with F. novicida showed no co-localization between F. novicida and caveolin-1. Arrowheads indicate the localization of F. novicida . Scale bar = 5m. (C) 5mM of amiloride was used to pre-treat BNL CL.2 cells and HeLa cells for 15 min prior to infection with F. novicida and Salmonella Thyphimurium (SL1344), respectively. Untreated cells were treated with media containing DMSO. Cells infected with F. novicida for 6 h followed by 2 h gentamicin treatment did not show a decrease in invasion while cells infected with S. Typhimurium for 30 min followed by 1 h gentamicin treatment showed a significant decrease in invasion *P
    Figure Legend Snippet: F. novicida entry does not utilize caveolin-dependent or macropinocytosis pathways. (A) BNL CL.2 and NMuLi cells were treated with the caveolin-inhibitor filipin and infected with F. novicida for 22 h followed by 2 h gentamicin treatment for invasion assays. (B) Immunolocalization of caveolin-1 (green) and DAPI (blue) of uninfected BNL CL.2 cells or following 8 h, 16 h, and 24 h infections with F. novicida showed no co-localization between F. novicida and caveolin-1. Arrowheads indicate the localization of F. novicida . Scale bar = 5m. (C) 5mM of amiloride was used to pre-treat BNL CL.2 cells and HeLa cells for 15 min prior to infection with F. novicida and Salmonella Thyphimurium (SL1344), respectively. Untreated cells were treated with media containing DMSO. Cells infected with F. novicida for 6 h followed by 2 h gentamicin treatment did not show a decrease in invasion while cells infected with S. Typhimurium for 30 min followed by 1 h gentamicin treatment showed a significant decrease in invasion *P

    Techniques Used: Infection

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    ATCC nmuli cells
    <t>miR-669c-3p</t> downregulates glutathione S-transferase (GST) protein expression. (A, B) Expression of GST mRNA (A) and protein (B) in <t>NMuLi</t> cells transfected with 20 nM miR-669c-3p mimic for 48 or 72 hr and evaluated by quantitative real-time PCR or Western blotting. Data are presented as the mean ± SEM ( n =4). *p
    Nmuli Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    miR-669c-3p downregulates glutathione S-transferase (GST) protein expression. (A, B) Expression of GST mRNA (A) and protein (B) in NMuLi cells transfected with 20 nM miR-669c-3p mimic for 48 or 72 hr and evaluated by quantitative real-time PCR or Western blotting. Data are presented as the mean ± SEM ( n =4). *p

    Journal: Bioscience of Microbiota, Food and Health

    Article Title: Sesame lignans upregulate glutathione S-transferase expression and downregulate microRNA-669c-3p

    doi: 10.12938/bmfh.2021-067

    Figure Lengend Snippet: miR-669c-3p downregulates glutathione S-transferase (GST) protein expression. (A, B) Expression of GST mRNA (A) and protein (B) in NMuLi cells transfected with 20 nM miR-669c-3p mimic for 48 or 72 hr and evaluated by quantitative real-time PCR or Western blotting. Data are presented as the mean ± SEM ( n =4). *p

    Article Snippet: Additionally, we assessed the effect of sesamin or episesamin on miR-669c-3p and GST expression in NMuLi cells.

    Techniques: Expressing, Transfection, Real-time Polymerase Chain Reaction, Western Blot

    (a) Expression of Gdf15 in parenchymal cells. Liver cells isolated from GalN-treated mice were separated into nonparenchymal and parenchymal fractions. HGF, hepatocyte growth factor. (b) Expression of Gdf15 in NMuLi and AML12 cells treated with CCl 4 in culture. Control flasks containing cells were sealed and incubated at 37°C in the absence of CCl 4 . Graphs to the right of the Northern blots show Gdf15 expression normalized to aldolase expression and plotted relative to uninduced levels. (c) Expression of Gdf15 following heat shock treatment of NMuLi and AML12 cells in culture. Cells were incubated at 44°C for 30 min starting at time zero and then allowed to recover at 37°C.

    Journal: Molecular and Cellular Biology

    Article Title: Characterization of Growth-Differentiation Factor 15, a Transforming Growth Factor ? Superfamily Member Induced following Liver Injury

    doi:

    Figure Lengend Snippet: (a) Expression of Gdf15 in parenchymal cells. Liver cells isolated from GalN-treated mice were separated into nonparenchymal and parenchymal fractions. HGF, hepatocyte growth factor. (b) Expression of Gdf15 in NMuLi and AML12 cells treated with CCl 4 in culture. Control flasks containing cells were sealed and incubated at 37°C in the absence of CCl 4 . Graphs to the right of the Northern blots show Gdf15 expression normalized to aldolase expression and plotted relative to uninduced levels. (c) Expression of Gdf15 following heat shock treatment of NMuLi and AML12 cells in culture. Cells were incubated at 44°C for 30 min starting at time zero and then allowed to recover at 37°C.

    Article Snippet: NMuLi cells (ATCC CRL-1638) and AML12 cells (ATCC CRL-2254) from the American Type Culture Collection (Manassas, Va.) were grown to near confluence in plastic flasks.

    Techniques: Expressing, Isolation, Mouse Assay, Incubation, Northern Blot

    Molecular analysis of cell clones. A) INDEL analysis of MnlI-TALEN and NcoI-TALEN N-Muli treated cells. The left and right arms of the TALEN pair are indicated in green and red, respectively. The restriction enzyme sites are indicated in blue. Deletions are indicated as dashes. The length of the deletions is indicated on the right of each sequence. B) Western blot analysis of the isolated cell clones. Proteins were prepared from the isolated cell clones and analyzed by Western blot using an anti-Ugt1 antibody. Lanes 1 and 2 correspond to protein extracts prepared from WT and mutant livers [13] . Lane 3 corresponds to untreated N-Muli cells. The arrow indicates the band corresponding to Ugt1. β-tubulin was used to normalize for protein loading. C) Ugt1 glucuronidation activity of the isolated cell clones. Ugt1 glucuronidation activity was determined by the UGT-Glo Assay as described in the Materials and Methods section. The activity determination was determined with microsomes prepared from wt and Ugt mutant mouse liver (1 µg), from wt N-Muli cells (10 µg) and from the isolated cell clones (10 µg, Mnl20, Nco20 and Nco25), as the percentage of luminescent substrate consumed. “+cntrl” indicates microsomes provided by the manufacturers (Promega) as positive control of the experiment. The experiment was repeated twice, and the mean of both experiments is shown.

    Journal: PLoS ONE

    Article Title: Generation of Ugt1-Deficient Murine Liver Cell Lines Using TALEN Technology

    doi: 10.1371/journal.pone.0104816

    Figure Lengend Snippet: Molecular analysis of cell clones. A) INDEL analysis of MnlI-TALEN and NcoI-TALEN N-Muli treated cells. The left and right arms of the TALEN pair are indicated in green and red, respectively. The restriction enzyme sites are indicated in blue. Deletions are indicated as dashes. The length of the deletions is indicated on the right of each sequence. B) Western blot analysis of the isolated cell clones. Proteins were prepared from the isolated cell clones and analyzed by Western blot using an anti-Ugt1 antibody. Lanes 1 and 2 correspond to protein extracts prepared from WT and mutant livers [13] . Lane 3 corresponds to untreated N-Muli cells. The arrow indicates the band corresponding to Ugt1. β-tubulin was used to normalize for protein loading. C) Ugt1 glucuronidation activity of the isolated cell clones. Ugt1 glucuronidation activity was determined by the UGT-Glo Assay as described in the Materials and Methods section. The activity determination was determined with microsomes prepared from wt and Ugt mutant mouse liver (1 µg), from wt N-Muli cells (10 µg) and from the isolated cell clones (10 µg, Mnl20, Nco20 and Nco25), as the percentage of luminescent substrate consumed. “+cntrl” indicates microsomes provided by the manufacturers (Promega) as positive control of the experiment. The experiment was repeated twice, and the mean of both experiments is shown.

    Article Snippet: Generation of mutant stable cell clones by TALEN transfection N-Muli cells (ATCC CRL-1638) were cultured in D-MEM with 10% FCS.

    Techniques: Sequencing, Western Blot, Isolation, Clone Assay, Mutagenesis, Activity Assay, Glo Assay, Positive Control

    Transient transfection of NcoI TALENs into N-Muli murine hepatoma cells. A) Determination of TALEN activity by RE-digestion of genomic PCR product. N-Muli cells were transfected with the NcoI TALEN pair together with a GFP plasmid. GFP positive cells were sorted, genomic DNA prepared and PCR amplified. PCR products were digested or not with NcoI, and the digestion product run in a 2% agarose gel. The arrow indicates the NcoI-resistant fraction of the PCR product to the NcoI-TALEN transfected cells (Lane 5). B) Confirmation of the presence of NcoI-resistant fragment. The agarose regions corresponding to the full-length undigested fragment of the gel shown in Panel A (GFP and NcoI-TALEN, lanes 4 and 5, respectively) were cut and DNA purified. A nested PCR reaction using internal primers was performed, and the PCR product was digested or not with NcoI (lanes 3, 6 and 2, 5, respectively). The percentage of uncut fragment is indicated bellow the gel. C) INDEL analysis of NcoI-TALEN N-Muli treated cells. The NcoI-resistant fragment of Panel A, Lane 5 was purified, cloned into a pUC19 vector and the obtained clones were sequenced. The left and right arms of the TALEN pair are indicated in green and red, respectively. The NcoI restriction enzyme site is indicated in blue. The deletions are indicated as black dashes. The type and length of the modification are indicated on the right of each sequence (Δ indicates deletion, and + base substitution, indicated by small orange caps).

    Journal: PLoS ONE

    Article Title: Generation of Ugt1-Deficient Murine Liver Cell Lines Using TALEN Technology

    doi: 10.1371/journal.pone.0104816

    Figure Lengend Snippet: Transient transfection of NcoI TALENs into N-Muli murine hepatoma cells. A) Determination of TALEN activity by RE-digestion of genomic PCR product. N-Muli cells were transfected with the NcoI TALEN pair together with a GFP plasmid. GFP positive cells were sorted, genomic DNA prepared and PCR amplified. PCR products were digested or not with NcoI, and the digestion product run in a 2% agarose gel. The arrow indicates the NcoI-resistant fraction of the PCR product to the NcoI-TALEN transfected cells (Lane 5). B) Confirmation of the presence of NcoI-resistant fragment. The agarose regions corresponding to the full-length undigested fragment of the gel shown in Panel A (GFP and NcoI-TALEN, lanes 4 and 5, respectively) were cut and DNA purified. A nested PCR reaction using internal primers was performed, and the PCR product was digested or not with NcoI (lanes 3, 6 and 2, 5, respectively). The percentage of uncut fragment is indicated bellow the gel. C) INDEL analysis of NcoI-TALEN N-Muli treated cells. The NcoI-resistant fragment of Panel A, Lane 5 was purified, cloned into a pUC19 vector and the obtained clones were sequenced. The left and right arms of the TALEN pair are indicated in green and red, respectively. The NcoI restriction enzyme site is indicated in blue. The deletions are indicated as black dashes. The type and length of the modification are indicated on the right of each sequence (Δ indicates deletion, and + base substitution, indicated by small orange caps).

    Article Snippet: Generation of mutant stable cell clones by TALEN transfection N-Muli cells (ATCC CRL-1638) were cultured in D-MEM with 10% FCS.

    Techniques: Transfection, TALENs, Activity Assay, Polymerase Chain Reaction, Plasmid Preparation, Amplification, Agarose Gel Electrophoresis, Purification, Nested PCR, Clone Assay, Modification, Sequencing

    Analysis of N-Muli Ugt1-mutant cell clones. A) and B) Schemes of the PCR fragments showing the position of the PCR primers and restriction sites. The position of the MnlI and NcoI sites are indicated (Panels A and B, respectively), as well as the expected length of the digestion products. C) and D) Restriction enzyme assay to detect INDELs. DNA from antibiotic-resistant clones (Panels C and D, from MnlI and NcoI TALEN-transfected cells, respectively) was prepared, PCR amplified, digested or not with MnlI and NcoI and the products run in a 1% agarose gel. The expected and obtained gel bands are indicated with arrows. N-Muli corresponds to control DNA, Mnl4, Mnl20, Nco25 and Nco20 to antibiotic-resistant clones that were transfected with the respective TALEN pairs.

    Journal: PLoS ONE

    Article Title: Generation of Ugt1-Deficient Murine Liver Cell Lines Using TALEN Technology

    doi: 10.1371/journal.pone.0104816

    Figure Lengend Snippet: Analysis of N-Muli Ugt1-mutant cell clones. A) and B) Schemes of the PCR fragments showing the position of the PCR primers and restriction sites. The position of the MnlI and NcoI sites are indicated (Panels A and B, respectively), as well as the expected length of the digestion products. C) and D) Restriction enzyme assay to detect INDELs. DNA from antibiotic-resistant clones (Panels C and D, from MnlI and NcoI TALEN-transfected cells, respectively) was prepared, PCR amplified, digested or not with MnlI and NcoI and the products run in a 1% agarose gel. The expected and obtained gel bands are indicated with arrows. N-Muli corresponds to control DNA, Mnl4, Mnl20, Nco25 and Nco20 to antibiotic-resistant clones that were transfected with the respective TALEN pairs.

    Article Snippet: Generation of mutant stable cell clones by TALEN transfection N-Muli cells (ATCC CRL-1638) were cultured in D-MEM with 10% FCS.

    Techniques: Mutagenesis, Polymerase Chain Reaction, Enzymatic Assay, Clone Assay, Transfection, Amplification, Agarose Gel Electrophoresis