Journal: Biological Procedures Online

Article Title: Construction of a bacterial autoinducer detection system in mammalian cells

doi: 10.1251/bpo98

Figure Lengend Snippet: COS-1 cells expressing either of the indicated QscR-based proteins were fixed and QscR proteins were detected using an antiserum directed against the Myc epitope as described in materials and methods. The nuclei of the cells were stained with, 6’-diamidino-2-phenylindone (DAPI, blue image in central panel). The left and center images were merged to detect overlap between the signals.

Article Snippet: The monkey kidney COS-1 cell line was cultured in Dulbecco’s modified Eagle’s medium (BioWhittaker, Walkersville, MD) supplemented with 10% Cosmic Calf Serum (HyClone, Logan, UT).

Techniques: Expressing, Staining

Journal: Biological Procedures Online

Article Title: Construction of a bacterial autoinducer detection system in mammalian cells

doi: 10.1251/bpo98

Figure Lengend Snippet: Mammalian expression vectors encoding the indicated RhlR-based chimeric proteins were co-transfected into COS-1 cells with either the (lasBOX 1)2-Luc or (lasBOX 2)2-Luc reporter plasmids and luciferase values were measured. The activity detected in cells transfected with the (lasBOX 1)2-Luc plasmid alone was set to 1.0 and the activity measured for the other transfections were calculated relative to this value. All data points were performed in triplicate and data representative of three repetitions are shown.

Article Snippet: The monkey kidney COS-1 cell line was cultured in Dulbecco’s modified Eagle’s medium (BioWhittaker, Walkersville, MD) supplemented with 10% Cosmic Calf Serum (HyClone, Logan, UT).

Techniques: Expressing, Transfection, Luciferase, Activity Assay, Plasmid Preparation

Journal: Biological Procedures Online

Article Title: Construction of a bacterial autoinducer detection system in mammalian cells

doi: 10.1251/bpo98

Figure Lengend Snippet: COS-1 cells were cotransfected with expression vectors encoding the indicated RhlR-based chimeric proteins and the (lasBOX 1)2-Luc reporter plasmid in the absence or presence of C4-HSL for 40 hours. Luciferase values were measured and expressed relative to the activity of the reporter plasmid alone.

Article Snippet: The monkey kidney COS-1 cell line was cultured in Dulbecco’s modified Eagle’s medium (BioWhittaker, Walkersville, MD) supplemented with 10% Cosmic Calf Serum (HyClone, Logan, UT).

Techniques: Expressing, Plasmid Preparation, Luciferase, Activity Assay

Journal: Biological Procedures Online

Article Title: Construction of a bacterial autoinducer detection system in mammalian cells

doi: 10.1251/bpo98

Figure Lengend Snippet: COS-1 cells were cotransfected with a mammalian expression plasmid encoding T3N-LasR and the (lasBOX 1)2-Luc reporter plasmid. The cells were incubated in the absence or presence of either 250μM 3O-C12-HSL or 750μM C12-HSL for three different time periods and luciferase activities were measured. The data are presented relative to the activity of the reporter plasmid alone. The highest activity of T3N-LasR was observed after 8 hours incubation with either 3O-C12-HSL or C12-HSL.

Article Snippet: The monkey kidney COS-1 cell line was cultured in Dulbecco’s modified Eagle’s medium (BioWhittaker, Walkersville, MD) supplemented with 10% Cosmic Calf Serum (HyClone, Logan, UT).

Techniques: Expressing, Plasmid Preparation, Incubation, Luciferase, Activity Assay