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pcr amplifying sec61β  (Addgene inc)
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Addgene inc pcr amplifying sec61β
BAF controls rupture diffusion in a size-dependent manner. A) Measurements of the width of the NE rupture gap in BJ5ta cells expressing <t>GFP-Sec61β.</t> Number of cells analyzed: siControl, n=15; siBAF, n=14. Error bars indicate ± SEM from triplicate experiments. B) Representative images of BJ5ta cells expressing either Hsp90-GFP or α-Tubulin-GFP after laser-induced NE rupture. Scale bar; 10 µm. C) Quantification of the nuclear-to-cytoplasmic ratio of cells expressing either Hsp90-GFP or α-tubulin-GFP treated with either siControl (n = 29 and 14, respectively) or siBAF (n = 21 and 16, respectively) from triplicate experiments. Error bars indicate ± SEM. D) Initial rate of increase into the nucleus following NE rupture for cells in B. E) Representative images of BJ5ta cells expressing Hsp90-GFP or α-tubulin-GFP during FLIP. Green circle indicates area of photobleaching in the cytoplasm. Red circle indicates area of measurement. F) FLIP measurements showing relative mobility of Hsp90-GFP (n = 16 cells) or α-tubulin-GFP (n = 16 cells). The slope of each line x was used to factor relative mobility between Hsp90-GFP and α-tubulin-GFP. G) Mobility of Hsp90-GFP and α-tubulin-GFP (measured as the best line-of-fit slope for each individual cell). H) Initial rate of increase into the nucleus following NE rupture for cells in D normalized by mobility. Error bars indicate ± SEM. Statistical significance: *, P<0.05; **<0.005; ***, P<0.0005 using an unpaired student t-test.
Pcr Amplifying Sec61β, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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BAF controls rupture diffusion in a size-dependent manner. A) Measurements of the width of the NE rupture gap in BJ5ta cells expressing GFP-Sec61β. Number of cells analyzed: siControl, n=15; siBAF, n=14. Error bars indicate ± SEM from triplicate experiments. B) Representative images of BJ5ta cells expressing either Hsp90-GFP or α-Tubulin-GFP after laser-induced NE rupture. Scale bar; 10 µm. C) Quantification of the nuclear-to-cytoplasmic ratio of cells expressing either Hsp90-GFP or α-tubulin-GFP treated with either siControl (n = 29 and 14, respectively) or siBAF (n = 21 and 16, respectively) from triplicate experiments. Error bars indicate ± SEM. D) Initial rate of increase into the nucleus following NE rupture for cells in B. E) Representative images of BJ5ta cells expressing Hsp90-GFP or α-tubulin-GFP during FLIP. Green circle indicates area of photobleaching in the cytoplasm. Red circle indicates area of measurement. F) FLIP measurements showing relative mobility of Hsp90-GFP (n = 16 cells) or α-tubulin-GFP (n = 16 cells). The slope of each line x was used to factor relative mobility between Hsp90-GFP and α-tubulin-GFP. G) Mobility of Hsp90-GFP and α-tubulin-GFP (measured as the best line-of-fit slope for each individual cell). H) Initial rate of increase into the nucleus following NE rupture for cells in D normalized by mobility. Error bars indicate ± SEM. Statistical significance: *, P<0.05; **<0.005; ***, P<0.0005 using an unpaired student t-test.

Journal: bioRxiv

Article Title: Mechanisms by which barrier-to-autointegration factor regulates dynamics of nucleocytoplasmic leakage and membrane repair following nuclear envelope rupture

doi: 10.1101/2023.12.21.572811

Figure Lengend Snippet: BAF controls rupture diffusion in a size-dependent manner. A) Measurements of the width of the NE rupture gap in BJ5ta cells expressing GFP-Sec61β. Number of cells analyzed: siControl, n=15; siBAF, n=14. Error bars indicate ± SEM from triplicate experiments. B) Representative images of BJ5ta cells expressing either Hsp90-GFP or α-Tubulin-GFP after laser-induced NE rupture. Scale bar; 10 µm. C) Quantification of the nuclear-to-cytoplasmic ratio of cells expressing either Hsp90-GFP or α-tubulin-GFP treated with either siControl (n = 29 and 14, respectively) or siBAF (n = 21 and 16, respectively) from triplicate experiments. Error bars indicate ± SEM. D) Initial rate of increase into the nucleus following NE rupture for cells in B. E) Representative images of BJ5ta cells expressing Hsp90-GFP or α-tubulin-GFP during FLIP. Green circle indicates area of photobleaching in the cytoplasm. Red circle indicates area of measurement. F) FLIP measurements showing relative mobility of Hsp90-GFP (n = 16 cells) or α-tubulin-GFP (n = 16 cells). The slope of each line x was used to factor relative mobility between Hsp90-GFP and α-tubulin-GFP. G) Mobility of Hsp90-GFP and α-tubulin-GFP (measured as the best line-of-fit slope for each individual cell). H) Initial rate of increase into the nucleus following NE rupture for cells in D normalized by mobility. Error bars indicate ± SEM. Statistical significance: *, P<0.05; **<0.005; ***, P<0.0005 using an unpaired student t-test.

Article Snippet: GFP-Sec61β in pBabe puro (a generous gift from Indra Chandrasekar) was created by PCR amplifying Sec61β (a gift from Gia Voeltz, Addgene plasmid #49154) and recombining into XhoI-SalI cut GFP pBabe puro.

Techniques: Diffusion-based Assay, Expressing