2016 hela  (ATCC)

Journal: EMBO Molecular Medicine

Article Title: ENOblock synergizes with colistin to treat Acinetobacter baumannii infections

doi: 10.1038/s44321-025-00331-2

Figure Lengend Snippet: ( A ) Time-kill curves of E. coli Ec MCR1 + and Kp10 strains in the presence of 2xMIC ENOblock for 24 h. ( B ) Membrane permeabilization of E. coli Ec MCR1 + and Kp10 strains in the presence of 0.5xMIC ENOblock, incubated for 10 min, was quantified by Typhon Scanner. Data are represented as mean ± SEM from three independent replicates and experiments. * P = 0.001: Ec MCR1 + vs Ec MCR1 + + ENOblock(two-tailed Student’s t test). ( C ) Analysis of E. coli Ec MCR1 + and Kp10 strains adhesion into HeLa cells with (1xMIC) and without ENOblock treatment. The data are presented as means ± SEM, * P = 0.015: Ec MCR1 + vs Ec MCR1 + + ENOblock and * P = 0.02: Kp10 vs Kp10 + ENOblock (two-tailed Student’s t test). Analysis of E. coli Ec MCR1 + and Kp10 strains invasion into HeLa cells with (1xMIC) and without ENOblock treatment. The data are presented as means ± SEM, * P = 0.002: Ec MCR1 + vs Ec MCR1 + + ENOblock and * P = 0.037: Kp10 vs Kp10 + ENOblock treatment (two-tailed Student’s t test).

Article Snippet: HeLa Cell Line , LGC Standards , Cat# ATCC-CCL-2.

Techniques: Membrane, Incubation, Two Tailed Test

Journal: EMBO Molecular Medicine

Article Title: ENOblock synergizes with colistin to treat Acinetobacter baumannii infections

doi: 10.1038/s44321-025-00331-2

Figure Lengend Snippet: ( A ) Schematic of the bacterial adhesion/invasion assay. ( B ) Analysis of Ab ATCC 17978 and Ab CR17 strains adhesion into HeLa and macrophage cells with (1xMIC) and without ENOblock treatment. The data are presented as means of three biological replicates ± SEM, For HeLa cells, * P = 0.023: Ab ATCC 17978 vs Ab ATCC 17978 + ENOblock and * P = 0.048: Ab CR17 vs Ab CR17 + ENOblock (two-tailed Student’s t test). For macrophage cells, * P = 0.018: Ab ATCC 17978 vs Ab ATCC 17978 + ENOblock and * P = 0.035: Ab CR17 vs Ab CR17 + ENOblock (two-tailed Student’s t test). ( C ) Analysis of Ab ATCC 17978 and Ab CR17 strains invasion into HeLa and macrophage cells with (1×MIC) and without ENOblock treatment. The data are presented as means of three biological replicates ± SEM, For HeLa cells, * P = 0.001: Ab ATCC 17978 vs Ab ATCC 17978 + ENOblock and * P = 0.011: Ab CR17 vs Ab CR17 + ENOblock (two-tailed Student’s t test). For macrophage cells, * P = 0.001: Ab ATCC 17978 vs Ab ATCC 17978 + ENOblock and * P = 0.002: Ab CR17 vs Ab CR17 + ENOblock (two-tailed Student’s t test). ( D ) Immunostaining of fibronectin of HeLa cells (magenta) and Ab ATCC 17978 and Ab CR17 strains (green) pretreated with ENOblock (0× and 1×MIC), after bacterial adherence for 2 h, was performed by specific primary antibodies against both strains and their respective secondary antibodies. Blue staining shows the location of HeLa cell nuclei. A representative image out of three biological replicates is shown. .

Article Snippet: HeLa Cell Line , LGC Standards , Cat# ATCC-CCL-2.

Techniques: Invasion Assay, Two Tailed Test, Immunostaining, Staining

Journal: EMBO Molecular Medicine

Article Title: ENOblock synergizes with colistin to treat Acinetobacter baumannii infections

doi: 10.1038/s44321-025-00331-2

Figure Lengend Snippet: ( A ) PCA plot of TMT data from Ab infected macrophages and epithelial cells of three biological replicates each group. ( B ) Venn diagrams showing the overlap of DEGs between the two comparisons: Ab infected macrophages vs Ab infected epithelial cells. The upper panel shows overlaps of overexpressed proteins, while the lower panel shows overlaps of downexpressed proteins. A total of six proteins were found to be commonly overexpressed in Ab upon exposure to both epithelial and macrophage cells: enolase, tyrosine–tRNA ligase, malonyl-CoA:acyl carrier protein transacylase, N-succinylarginine dihydrolase, carbonic anhydrase, and biotin carboxylase. Additionally, one protein, isocitrate dehydrogenase, was consistently downregulated in response to both cell types. ( C ) COG category distribution of DEPs. The overexpressed and downexpressed proteins in each comparison are categorized by their COG functional groups, with blue representing downexpressed proteins and orange representing overexpressed proteins. ( D ) Volcano plots depicting the DEPs for the two comparisons. Proteins significantly overexpressed are shown in red, while significantly downexpressed proteins are shown in blue. Enolase overexpression is shown in green. Non-significant proteins are indicated in black. Three biological replicates from each group were compared. ( E ) A. baumannii Ab ATCC 17978 and Δ eno strains, with or without supplementation of 4 mM phosphoenolpyruvate, adhesion to HeLa cells. The data are presented as means of three biological replicates ± SEM, and Student t test was used for statistical analysis. * P = 0.006: Ab ATCC 17978 vs Δ eno and * P < 0.001: Ab ATCC 17978 vs Δ eno + PEP (two-tailed Student’s t test). MΦ macrophage cells, Epit epithelial cells. .

Article Snippet: HeLa Cell Line , LGC Standards , Cat# ATCC-CCL-2.

Techniques: Infection, Comparison, Functional Assay, Over Expression, Two Tailed Test

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