parent cell line  (ATCC)

Journal: PLOS ONE

Article Title: Visualizing VDAC1 in live cells using a tetracysteine tag

doi: 10.1371/journal.pone.0311107

Figure Lengend Snippet: ( A ) AlphaFold2-generated 3D structures and schematic representation of GFP-VDAC1 and VDAC1-GFP fusion constructs. ( B ) Confocal images of HeLa cells stained with mitotracker red and transfected with GFP-VDAC1 (top panel) or VDAC1-GFP (bottom panel). Bar graphs (right) show the percentage of HeLa cells with different expression phenotypes of GFP-VDAC1 (top) or VDAC1-GFP (bottom) using different amounts of plasmid DNA.

Article Snippet: HeLa S3 cells, a clonal derivative of the parent HeLa cell line, were obtained from ATCC and authenticated with STR analysis.

Techniques: Generated, Construct, Staining, Transfection, Expressing, Plasmid Preparation

Journal: PLOS ONE

Article Title: Visualizing VDAC1 in live cells using a tetracysteine tag

doi: 10.1371/journal.pone.0311107

Figure Lengend Snippet: ( A ) 3D structure of VDAC1-TC and chemical structure of FlAsH (top). Confocal images of HeLa cells transfected with VDAC1-TC and mitoDsRed; cells were labeled with 1 μM FlAsH for 15 min and washed with 100 μM BAL for 10 min (bottom). Left image shows an overview of transfected and untransfected cells, and the numbered dashed squares are magnified on the right. ( B ) Immunofluorescence images of a HeLa cell expressing VDAC1-TC; cell was labeled with 1 μM FlAsH for 15 min (green) and anti-VDAC1 (magenta). Graph on the right shows the Pearson correlation between VDAC1-TC and Anti-VDAC1 signal. ( C ) 3D structure of VDAC1-TC and chemical structure of ReAsH (left). Confocal images of a HeLa cell expressing VDAC1-TC and Hexokinase1-GFP; cells were labeled with 166 nM ReAsH for 15 min and washed with 100 μM BAL for 10 min (right). Dashed lines represent the line scan graph on the right and show the relative fluorescence intensity of Hexokinase1-GFP (green) and VDAC1-TC (magenta) along the length of the line.

Article Snippet: HeLa S3 cells, a clonal derivative of the parent HeLa cell line, were obtained from ATCC and authenticated with STR analysis.

Techniques: Transfection, Labeling, Immunofluorescence, Expressing, Fluorescence

Journal: PLOS ONE

Article Title: Visualizing VDAC1 in live cells using a tetracysteine tag

doi: 10.1371/journal.pone.0311107

Figure Lengend Snippet: ( A ) Confocal images of a HeLa cell expressing VDAC1-TC, mitoCFP, and mCh-ER3; the cell was labeled with 1 μM FlAsH for 15 min and washed with 100 μM Bal for 10 min. The left image shows an overview of the cell, and the dashed squares are magnified on the right side. Yellow arrows point to positions of VDAC1-clusters at ER-mitochondria contact sites. Dashed lines represent the line scan graphs on the right and show the relative fluorescence intensity of VDAC1-TC (green), mitoCFP (blue), and mCh-ER3 (magenta) along the length of the line. ( B ) The graph shows the percentage of VDAC1-clusters that colocalize with the ER (n = 8). ( C ) Structured illumination microscopy images of a HeLa cell expressing VDAC1-TC with mCh-Sec61β; cell was labeled with 1 μM FlAsH for 15 min and washed with 100 μM BAL for 10 min. Dashed line represent the line scan graph on the right and show the relative fluorescence intensity of VDAC1-TC (green) and mCh-Sec61β (magenta) along the length of the line. Arrow point to the position of a VDAC1-cluster at ER contact sites.

Article Snippet: HeLa S3 cells, a clonal derivative of the parent HeLa cell line, were obtained from ATCC and authenticated with STR analysis.

Techniques: Expressing, Labeling, Fluorescence, Microscopy

Journal: PLOS ONE

Article Title: Visualizing VDAC1 in live cells using a tetracysteine tag

doi: 10.1371/journal.pone.0311107

Figure Lengend Snippet: Confocal images of a HeLa cell expressing VDAC1-TC and GFP-BAK with 10 mM glucose (left) and after 20 min of glucose depletion (middle); the cell was labeled with 333 nM ReAsH for 15 min. The first row shows an overview of the cell, and the dashed squares are magnified below. White arrows point to the contact site between VDAC1- and BAK-clusters. The graph shows the percentage of VDAC1-clusters that colocalized with BAK-clusters and the percentage of BAK-clusters that colocalized with VDAC1-clusters after 20 min of glucose depletion (right, n = 8 cells).

Article Snippet: HeLa S3 cells, a clonal derivative of the parent HeLa cell line, were obtained from ATCC and authenticated with STR analysis.

Techniques: Expressing, Labeling

Journal: PLOS ONE

Article Title: Visualizing VDAC1 in live cells using a tetracysteine tag

doi: 10.1371/journal.pone.0311107

Figure Lengend Snippet: ( A ) Confocal images of HeLa cells expressing mitoDsRed (top row) or VDAC1-TC with mitoDsRed (bottom row); cells were labeled with 1 μM FlAsH for 15 min and washed with 100 μM BAL for 10 min. The left column shows an overview of the cell, the middle column shows the Otsu thresholded mitoDsRed signal, and the dashed squares are magnified in the right column. Graphs on the right show the Aspect Ratio and Form Factor of thresholded images (n = 32 per group). The difference between groups was evaluated using unpaired t-test. Data are presented as mean ± SD. ( B ) Confocal images of a HeLa cells expressing VDAC1-TC and mitoDsRed; the cell was labeled with 1 μM FlAsH for 15 min and washed with 100 μM BAL for 10 min. On the left panel, an overview of the cell is shown, and the dashed square is magnified on the right side before and 10 min after perfusion with 2 μM FCCP. White arrows point to a VDAC1-cluster before and after mitochondrial fission. ( C ) Confocal images of a HeLa cell expressing VDAC1-TC and mitoDsRed; the cell was labeled with 1 μM FlAsH for 15 min and washed with 100 μM BAL for 10 min. On the right panel, an overview of the cell is shown, and the dashed square is magnified on the right side before and 10 min after perfusion with 4 μM Ionomycin. White arrows point to a VDAC1-cluster before and after mitochondrial fission.

Article Snippet: HeLa S3 cells, a clonal derivative of the parent HeLa cell line, were obtained from ATCC and authenticated with STR analysis.

Techniques: Expressing, Labeling