transfected construct (human (Microsynth ag)

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Microsynth ag transfected construct (human

( A ) Vectors encoding the indicated inner nuclear membrane (INM) proteins were <t>transfected</t> into HeLa cells. Cells were fixed after 24 or 48 hr and analyzed by confocal microscopy. Scale bar, 10 μm. ( B ) Time-lapse images of LAP1B-GFP or LAP2β-GFP expressing HeLa cells progressing through mitosis. Expression of the constructs was induced 24 hr prior to imaging. DNA was visualized by SirHoechst and used to define anaphase onset (t = 00:00 hr:min). Scale bar, 5 μm. ( C ) Scheme depicting the two human LAP1 isoforms, LAP1B and LAP1C. ( D ) Representative images of stable HeLa cells lines expressing LAP1B-GFP and LAP1C-GFP after induction with different tetracycline (tet) concentrations for 48 hr. Please note that we observed some ‘twin’ nuclei upon overexpression of LAP1B (white arrow), which can be a sign of binucleation, as further analyzed in . Scale bar, 10 μm. ( E ) LAP1 levels at the NE of cells from the experiment shown in panel D were analyzed by quantification of the integrated GFP density (IntDen GFP) per nucleus. ( F ) LAP1 levels in cell lysates derived from the experiment shown in panel D were analyzed by immunoblotting. Note that the LAP1B-GFP cell line expressed LAP1C-GFP independent of tetracycline addition, due to an alternative transcriptional start site used for the production of the shorter LAP1 isoform . ( G ) Left: Maximum intensity z-projections (5 × 0.63 μm) of confocal images from fixed metaphase HeLa cells expressing LAP1B-GFP or LAP1C-GFP after 48 hr of induction. Scale bar, 5 μm. Right: Confocal images of fixed HeLa cells in interphase. Scale bar, 10 μm.

Transfected Construct (Human, supplied by Microsynth ag, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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transfected construct (human - by Bioz Stars, 2026-11

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Images

1) Product Images from "Torsin ATPases influence chromatin interaction of the Torsin regulator LAP1"

Article Title: Torsin ATPases influence chromatin interaction of the Torsin regulator LAP1

Journal: eLife

doi: 10.7554/eLife.63614

( A ) Vectors encoding the indicated inner nuclear membrane (INM) proteins were transfected into HeLa cells. Cells were fixed after 24 or 48 hr and analyzed by confocal microscopy. Scale bar, 10 μm. ( B ) Time-lapse images of LAP1B-GFP or LAP2β-GFP expressing HeLa cells progressing through mitosis. Expression of the constructs was induced 24 hr prior to imaging. DNA was visualized by SirHoechst and used to define anaphase onset (t = 00:00 hr:min). Scale bar, 5 μm. ( C ) Scheme depicting the two human LAP1 isoforms, LAP1B and LAP1C. ( D ) Representative images of stable HeLa cells lines expressing LAP1B-GFP and LAP1C-GFP after induction with different tetracycline (tet) concentrations for 48 hr. Please note that we observed some ‘twin’ nuclei upon overexpression of LAP1B (white arrow), which can be a sign of binucleation, as further analyzed in . Scale bar, 10 μm. ( E ) LAP1 levels at the NE of cells from the experiment shown in panel D were analyzed by quantification of the integrated GFP density (IntDen GFP) per nucleus. ( F ) LAP1 levels in cell lysates derived from the experiment shown in panel D were analyzed by immunoblotting. Note that the LAP1B-GFP cell line expressed LAP1C-GFP independent of tetracycline addition, due to an alternative transcriptional start site used for the production of the shorter LAP1 isoform . ( G ) Left: Maximum intensity z-projections (5 × 0.63 μm) of confocal images from fixed metaphase HeLa cells expressing LAP1B-GFP or LAP1C-GFP after 48 hr of induction. Scale bar, 5 μm. Right: Confocal images of fixed HeLa cells in interphase. Scale bar, 10 μm.

Figure Legend Snippet: ( A ) Vectors encoding the indicated inner nuclear membrane (INM) proteins were transfected into HeLa cells. Cells were fixed after 24 or 48 hr and analyzed by confocal microscopy. Scale bar, 10 μm. ( B ) Time-lapse images of LAP1B-GFP or LAP2β-GFP expressing HeLa cells progressing through mitosis. Expression of the constructs was induced 24 hr prior to imaging. DNA was visualized by SirHoechst and used to define anaphase onset (t = 00:00 hr:min). Scale bar, 5 μm. ( C ) Scheme depicting the two human LAP1 isoforms, LAP1B and LAP1C. ( D ) Representative images of stable HeLa cells lines expressing LAP1B-GFP and LAP1C-GFP after induction with different tetracycline (tet) concentrations for 48 hr. Please note that we observed some ‘twin’ nuclei upon overexpression of LAP1B (white arrow), which can be a sign of binucleation, as further analyzed in . Scale bar, 10 μm. ( E ) LAP1 levels at the NE of cells from the experiment shown in panel D were analyzed by quantification of the integrated GFP density (IntDen GFP) per nucleus. ( F ) LAP1 levels in cell lysates derived from the experiment shown in panel D were analyzed by immunoblotting. Note that the LAP1B-GFP cell line expressed LAP1C-GFP independent of tetracycline addition, due to an alternative transcriptional start site used for the production of the shorter LAP1 isoform . ( G ) Left: Maximum intensity z-projections (5 × 0.63 μm) of confocal images from fixed metaphase HeLa cells expressing LAP1B-GFP or LAP1C-GFP after 48 hr of induction. Scale bar, 5 μm. Right: Confocal images of fixed HeLa cells in interphase. Scale bar, 10 μm.

Techniques Used: Transfection, Confocal Microscopy, Expressing, Construct, Imaging, Over Expression, Derivative Assay, Western Blot

( A ) Vector encoding LAP1B-GFP was transiently transfected into HCT116 or HepG2 cells. Cells were fixed after 48 hr and analyzed by confocal microscopy. Scale bars, 10 μm. ( B ) Western blot analysis (left) and integrated GFP fluorescence intensities (right) in LAP1B-GFP and LAP2β-GFP-expressing cells of the experiment depicted in . ( C ) Comparison of the metaphase localization of nuclear lamins and select inner nuclear membrane (INM) proteins in an inducible LAP1B-GFP cell line without and with tetracycline (tet) induction. Maximum intensity projections (5 × 0.63 μm). Scale bar, 5 μm.

Figure Legend Snippet: ( A ) Vector encoding LAP1B-GFP was transiently transfected into HCT116 or HepG2 cells. Cells were fixed after 48 hr and analyzed by confocal microscopy. Scale bars, 10 μm. ( B ) Western blot analysis (left) and integrated GFP fluorescence intensities (right) in LAP1B-GFP and LAP2β-GFP-expressing cells of the experiment depicted in . ( C ) Comparison of the metaphase localization of nuclear lamins and select inner nuclear membrane (INM) proteins in an inducible LAP1B-GFP cell line without and with tetracycline (tet) induction. Maximum intensity projections (5 × 0.63 μm). Scale bar, 5 μm.

Techniques Used: Plasmid Preparation, Transfection, Confocal Microscopy, Western Blot, Fluorescence, Expressing

$( A ) HeLa cells were transiently transfected with constructs encoding for LAP1B-GFP or LAP1B(1-359), either alone or together with the depicted Torsin constructs. After 48 hr, cells were fixed, subjected to immunostaining of Torsins using an anti-HA antibody and analyzed by confocal microscopy. Scale bar, 10 μm. ( B ) Representative images of pre-defined classes of cell nuclei used for the classification of NE aberration phenotypes. Based on nuclear shape, cells were assigned into one of the five shown categories for the quantification in C. The two upper classes were regarded as normal (no; no NE aberration), and the three lower classes as aberrant (yes). Scale bar, 5 μm. ( C ) The percentage of cells with NE aberrations was quantified by assigning cells into one of five predefined phenotypic classes shown in B. (left; N = 3, n > 103, mean +/- SEM). Only cells with similar LAP1B-GFP expression levels, based on quantification of the integrated GFP density per cell (IntDen GFP, normalized to the LAP1B minus Torsin control; right) were considered. ( D ) Depiction of the mutant LAP1B constructs; R563G: Torsin activation deficient-mutant; E482A, disease-causing mutant. ( E ) HeLa cells were transiently transfected with constructs encoding for the indicated LAP1B-GFP derivatives, either alone or together with Tor1B-mRFP. After 48 hr, cells were fixed and analyzed by confocal microscopy. Scale bar, 10 μm. ( F ) The fraction of cells with NE aberrations was quantified as in C (N = 3, n > 114, mean +/- SEM).$
Figure Legend Snippet: ( A ) HeLa cells were transiently transfected with constructs encoding for LAP1B-GFP or LAP1B(1-359), either alone or together with the depicted Torsin constructs. After 48 hr, cells were fixed, subjected to immunostaining of Torsins using an anti-HA antibody and analyzed by confocal microscopy. Scale bar, 10 μm. ( B ) Representative images of pre-defined classes of cell nuclei used for the classification of NE aberration phenotypes. Based on nuclear shape, cells were assigned into one of the five shown categories for the quantification in C. The two upper classes were regarded as normal (no; no NE aberration), and the three lower classes as aberrant (yes). Scale bar, 5 μm. ( C ) The percentage of cells with NE aberrations was quantified by assigning cells into one of five predefined phenotypic classes shown in B. (left; N = 3, n > 103, mean +/- SEM). Only cells with similar LAP1B-GFP expression levels, based on quantification of the integrated GFP density per cell (IntDen GFP, normalized to the LAP1B minus Torsin control; right) were considered. ( D ) Depiction of the mutant LAP1B constructs; R563G: Torsin activation deficient-mutant; E482A, disease-causing mutant. ( E ) HeLa cells were transiently transfected with constructs encoding for the indicated LAP1B-GFP derivatives, either alone or together with Tor1B-mRFP. After 48 hr, cells were fixed and analyzed by confocal microscopy. Scale bar, 10 μm. ( F ) The fraction of cells with NE aberrations was quantified as in C (N = 3, n > 114, mean +/- SEM).

Techniques Used: Transfection, Construct, Immunostaining, Confocal Microscopy, Expressing, Mutagenesis, Activation Assay

( A ) HeLa cells were transfected with constructs encoding for LAP1B-GFP, either alone or together with LULL1-HASt and Tor1B-mRFP expression vectors. Cell were fixed 48 hr after transfection and nuclear morphology analyzed by confocal microscopy. Scale bar, 10 μm. ( B ) Quantification of NE aberrations in cells with similar LAP1B-GFP expression levels as in (N = 3, n > 99, mean +/- SEM).

Figure Legend Snippet: ( A ) HeLa cells were transfected with constructs encoding for LAP1B-GFP, either alone or together with LULL1-HASt and Tor1B-mRFP expression vectors. Cell were fixed 48 hr after transfection and nuclear morphology analyzed by confocal microscopy. Scale bar, 10 μm. ( B ) Quantification of NE aberrations in cells with similar LAP1B-GFP expression levels as in (N = 3, n > 99, mean +/- SEM).

Techniques Used: Transfection, Construct, Expressing, Confocal Microscopy

Figure Legend Snippet:

Techniques Used: Transfection, Construct, Recombinant, Molecular Cloning, Knock-Out

Transfection:

Article Title: Torsin ATPases influence chromatin interaction of the Torsin regulator LAP1
Article Snippet: Transfected construct (human) , si-Tor1B , Qiagen/Microsynth , , 5’- CTGTCGGAGTCTTCAATAATA-3’.

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transfected construct (human (Microsynth ag)

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1) Product Images from "Torsin ATPases influence chromatin interaction of the Torsin regulator LAP1"

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