Journal: Chemosensors

Article Title: Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis

doi: 10.3390/chemosensors12110223

Figure Lengend Snippet: Figure 2. The effect of overexpression of different proteins on the bioluminescence emission in mammalian cell lines: (A) LiveLight HEK293; (B) HEK293; (C) HeLa. Cells grown in 24-well plates were transfected with a mixture of 0.4 µg lux plasmids and 0.1 µg of the indicated genes (all constructs in pcDNA3.1(+)). The signal was normalized to the bioluminescence emission of cells transfected with 0.4 µg lux plasmids and 0.1 µg of the empty pcDNA3.1(+) vector (−). For LiveLight HEK293, 0.5 µg of the indicated gene in pcDNA3.1(+) was transfected. Error bars represent standard deviation from 5 wells.

Article Snippet: HeLa cells were obtained from LGC Standards (Wesel, Germany, cat. no. ATCC-CCL-2).

Techniques: Over Expression, Transfection, Construct, Plasmid Preparation, Standard Deviation

Journal: Chemosensors

Article Title: Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis

doi: 10.3390/chemosensors12110223

Figure Lengend Snippet: Figure 3. Fluorescence ratio of iNap1 in Lux-expressing cells with and without Akt2CA. (A) LiveLight HEK293; (B) HEK293; (C) HeLa cells grown on coverslips were transfected with a mixture of 0.6 µg lux plasmids, 0.2 µg iNap1 pcDNA3.1 Hygro(+) and 0.2 µg Akt2CA pcDNA3.1(+) or the empty pcDNA3.1(+) vector (–). For LiveLight HEK293, 0.2 µg iNap1 pcDNA3.1 Hygro(+) and 0.8 µg Akt2CA pcDNA3.1(+) or the empty pcDNA3.1(+) vector were transfected. iNap1 fluorescence excited at 405 (F405) and 491 nm (F491) was recorded with a custom-built microscope. Error bars represent standard deviation from at least 50 cells. ** and *** represent p values of <0.01 and <0.001, respectively, calculated by a 2-tailed Student’s t test.

Article Snippet: HeLa cells were obtained from LGC Standards (Wesel, Germany, cat. no. ATCC-CCL-2).

Techniques: Fluorescence, Expressing, Transfection, Plasmid Preparation, Microscopy, Standard Deviation

Journal: Chemosensors

Article Title: Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis

doi: 10.3390/chemosensors12110223

Figure Lengend Snippet: Figure 5. MTT assay of cells transfected with Akt2CA-P2A-RFK. (A) LiveLight HEK293; (B) HEK293; (C) HeLa cells grown in 24-well plates were transfected with a mixture of 0.1 µg Akt2CA-P2A-RFK and 0.4 µg of the empty vector or 0.5 µg of the empty vector (−) (all constructs in pcDNA3.1(+)). For LiveLight HEK293, 0.5 µg of the empty vector or the Akt2CA-P2A-RFK plasmid was transfected. Absorbance of cell lysates was measured at 570 nm. Error bars represent standard deviation from 10 wells.

Article Snippet: HeLa cells were obtained from LGC Standards (Wesel, Germany, cat. no. ATCC-CCL-2).

Techniques: MTT Assay, Transfection, Plasmid Preparation, Construct, Standard Deviation

Journal: Chemosensors

Article Title: Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis

doi: 10.3390/chemosensors12110223

Figure Lengend Snippet: Figure 4. Effect of combined expression of RFK and Akt2CA on the bioluminescence emission in different mammalian cell lines: (A) LiveLight HEK293; (B) HEK293; (C) HeLa. Cells were grown in 24-well plates and transfected with a mixture of 0.4 µg lux plasmids and 0.1 µg of the indicated constructs (all in pcDNA3.1(+)). For RFK + Akt2CA, two separate plasmids containing RFK and Akt2CA were cotransfected (0.05 µg each). The signal was normalized to cells transfected with 0.4 µg lux plasmids and 0.1 µg of the empty pcDNA3.1(+) vector (−). For LiveLight HEK293, 0.5 µg of the indicated constructs was transfected. Error bars represent standard deviation from 5 wells.

Article Snippet: HeLa cells were obtained from LGC Standards (Wesel, Germany, cat. no. ATCC-CCL-2).

Techniques: Expressing, Transfection, Construct, Plasmid Preparation, Standard Deviation

Journal: Chemosensors

Article Title: Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis

doi: 10.3390/chemosensors12110223

Figure Lengend Snippet: Figure 6. Bioluminescence of HeLa cells with and without Akt2CA-P2A-RFK. Cells grown on coverslips were transfected with a mixture of 0.8 µg lux plasmids and (A) 0.2 µg empty pcDNA3.1(+) vector or (B) 0.2 µg Akt2CA-P2A-RFK (all constructs in pcDNA3.1(+)). Bioluminescence emission was recorded using the indicated exposure times. The colormap was adjusted to the minimum and maximum pixel value of each image.

Article Snippet: HeLa cells were obtained from LGC Standards (Wesel, Germany, cat. no. ATCC-CCL-2).

Techniques: Transfection, Plasmid Preparation, Construct