l2 fibroblast cell line  (Thermo Fisher)

Journal: PLOS Pathogens

Article Title: Listeria monocytogenes requires phosphotransferase systems to facilitate intracellular growth and virulence

doi: 10.1371/journal.ppat.1012492

Figure Lengend Snippet: (A) Intracellular growth of WT, Δ glpD /Δ golD , Δ uhpT, Δ glpD /Δ golD /Δ uhpT was determined in BMDMs following infection at an MOI of 0.2. Growth curves are representative of at least three independent experiments. Error bars represent the standard deviation of the means of technical triplicates within the representative experiment. (B) L2 fibroblasts were infected with indicated L. monocytogenes strains at an MOI of 0.5 and were examined for plaque formation 4 days post infection. Assays were performed in biological triplicate and data displayed is the median and SEM of a strain’s plaque size relative to WT in one of three representative biological replicates. (C) Bacterial burdens from the spleen and liver were enumerated at 48 hours post-intravenous infection with 1x10 5 bacteria. Data are representative of results from two separate experiments. Horizontal dashed lines represent the limits of detection, and the bars associated with the individual strains represents the mean and SEM of the group.

Article Snippet: Plaque assays were conducted using a L2 fibroblast cell line grown in Dulbecco’s Minimal Essential Media (DMEM) based media (Thermo Fischer: 11965092) as previously described with minor modifications for visualization and quantification of plaques [ , ].

Techniques: Infection, Standard Deviation, Bacteria

Journal: PLOS Pathogens

Article Title: Listeria monocytogenes requires phosphotransferase systems to facilitate intracellular growth and virulence

doi: 10.1371/journal.ppat.1012492

Figure Lengend Snippet: (A) PTS mediated free sugar import and phosphorylation by phosphocarrier protein phospho-cycling from the terminal conversion of phosphoenol-pyruvate (PEP) to pyruvate. (B) Intracellular growth of WT, Δ glpD /Δ golD /Δ uhpT , Δ ptsI , and Δ glpD /Δ golD /Δ uhpT /Δ ptsI was determined in BMDMs following infection at an MOI of 0.2. Growth curves are representative of at least three independent experiments. Error bars represent the standard deviation of the means of technical triplicates within the representative experiment. (C) L2 fibroblasts were infected with indicated L. monocytogenes strains at an MOI of 0.5 and were examined for plaque formation 4 days post infection. Assays were performed in biological triplicate and data displayed is the median and SEM of a strain’s plaque size relative to WT in one of three representative biological replicates. (D) Bacterial burdens from the spleen and liver were enumerated at 48 hours post-intravenous infection with 1x10 5 bacteria. Data are representative of results from two separate experiments. Horizontal dashed lines represent the limits of detection, and the bars associated with the individual strains represents the mean and SEM of the group.

Article Snippet: Plaque assays were conducted using a L2 fibroblast cell line grown in Dulbecco’s Minimal Essential Media (DMEM) based media (Thermo Fischer: 11965092) as previously described with minor modifications for visualization and quantification of plaques [ , ].

Techniques: Phospho-proteomics, Infection, Standard Deviation, Bacteria

Journal:

Article Title: Inhibition of ROCK activity allows InlF-mediated invasion and increased virulence of Listeria monocytogenes

doi: 10.1111/j.1365-2958.2008.06188.x

Figure Lengend Snippet: Cell type specific effects of ROCK activity inhibition on L. monocytogenes infection. Monolayers of cell lines were treated with 10 μM Y27632 or DMSO (control) for 30 min prior to washing of monolayers and infection with wild-type L. monocytogenes 10403S. Cell-associated (total) and intracellular bacteria were quantified by gentamicin protection assay as described in Experimental Procedures. Data presented represents the means ±s.d. fold change in CFU upon Y27632 treatment relative to control samples for one of three experiments performed in triplicate with similar results. * p <0.01; bold type indicates changes in CFU greater than 2-fold with p< 0.05.

Article Snippet: The human-derived epithelial cell lines HeLa (ATCC CCL-2) and HEp-2 (ATCC CCL-23), the murine-derived epithelial cell line 1308.1, the murine-derived fibroblast cell lines L2 and L929, the murine-derived macrophage cell lines RAW 264.7 and RAW 309 Cr.1, and the human-derived macrophage cell line U937 (ATCC CRL-1593.2) were cultured in RPMI 1640 medium (Mediatech, Herndon, VA) supplemented with 10% FBS (HyClone, Logan, UT), 55 μM 2-β-mercaptoethanol, 1 mM sodium pyruvate, and 2 mM glutamine.

Techniques: Activity Assay, Inhibition, Infection, Control, Bacteria

Journal:

Article Title: Inhibition of ROCK activity allows InlF-mediated invasion and increased virulence of Listeria monocytogenes

doi: 10.1111/j.1365-2958.2008.06188.x

Figure Lengend Snippet: Cell type specific effects of ROCK activity inhibition on L. monocytogenes infection. Monolayers of cell lines were treated with 10 μM Y27632 or DMSO (control) for 30 min prior to washing of monolayers and infection with wild-type L. monocytogenes 10403S. Cell-associated (total) and intracellular bacteria were quantified by gentamicin protection assay as described in Experimental Procedures. Data presented represents the means ±s.d. fold change in CFU upon Y27632 treatment relative to control samples for one of three experiments performed in triplicate with similar results. * p <0.01; bold type indicates changes in CFU greater than 2-fold with p< 0.05.

Article Snippet: The human-derived epithelial cell lines HeLa (ATCC CCL-2) and HEp-2 (ATCC CCL-23), the murine-derived epithelial cell line 1308.1, the murine-derived fibroblast cell lines L2 and L929, the murine-derived macrophage cell lines RAW 264.7 and RAW 309 Cr.1, and the human-derived macrophage cell line U937 (ATCC CRL-1593.2) were cultured in RPMI 1640 medium (Mediatech, Herndon, VA) supplemented with 10% FBS (HyClone, Logan, UT), 55 μM 2-β-mercaptoethanol, 1 mM sodium pyruvate, and 2 mM glutamine.

Techniques: Activity Assay, Inhibition, Infection