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Studies on the morphology and spreading of human endothelial cells define key inter- and intramolecular interactions for talin1.

Kopp PM, Bate N, Hansen TM, Brindle NP, Praekelt U, Debrand E, Coleman S, Mazzeo D, Goult BT, Gingras AR, Pritchard CA, Critchley DR, Monkley SJ - Eur. J. Cell Biol. (2010)

Bottom Line: The talin rod contains several actin-binding sites (ABS), and mutations in the C-terminal ABS that reduced actin-binding impaired talin1 function, whereas those that increased binding resulted in more stable FAs.The results show that both the N-terminal integrin and C-terminal actin-binding functions of talin are essential to cell spreading and FA assembly.Finally, mutations that relieve talin auto-inhibition resulted in the rapid and excessive production of FA, highlighting the importance of talin regulation within the cell.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, University of Leicester, Leicester, UK.

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Analysis of MEFs for talin structure function studies. (A) Western blots of Tln1fl/fl; CreER/+ MEF lysates 72 h after treatment with 100 nM 4-hydroxy tamoxifen (4OHT) or ethanol (EtOH) probed with talin 1 and 2 specific antibodies with vinculin as a loading control. (B) Numbers of GFP- or paxillin-containing FA in Tln1fl/fl; CreER/+ MEF 72 h after treatment with 100 nM 4OHT and either untransfected or transfected as shown. Results are expressed as mean ± s.e.m. (C) Different batches of primary MEFs differ in their phenotype before and after treatment with 4OHT. (Top panels) Brightfield images of two different Tln1fl/fl; CreER/+ MEF lines 96 h after treatment with either ethanol (EtOH) or 100 nM 4OHT. (Bottom panels) Western blot of lysates of same MEF lines 24 h or 96 h after treatment with either ethanol (−) or 100 nM 4OHT (+). Vinculin was used as a loading control.
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d32e1464: Analysis of MEFs for talin structure function studies. (A) Western blots of Tln1fl/fl; CreER/+ MEF lysates 72 h after treatment with 100 nM 4-hydroxy tamoxifen (4OHT) or ethanol (EtOH) probed with talin 1 and 2 specific antibodies with vinculin as a loading control. (B) Numbers of GFP- or paxillin-containing FA in Tln1fl/fl; CreER/+ MEF 72 h after treatment with 100 nM 4OHT and either untransfected or transfected as shown. Results are expressed as mean ± s.e.m. (C) Different batches of primary MEFs differ in their phenotype before and after treatment with 4OHT. (Top panels) Brightfield images of two different Tln1fl/fl; CreER/+ MEF lines 96 h after treatment with either ethanol (EtOH) or 100 nM 4OHT. (Bottom panels) Western blot of lysates of same MEF lines 24 h or 96 h after treatment with either ethanol (−) or 100 nM 4OHT (+). Vinculin was used as a loading control.


Studies on the morphology and spreading of human endothelial cells define key inter- and intramolecular interactions for talin1.

Kopp PM, Bate N, Hansen TM, Brindle NP, Praekelt U, Debrand E, Coleman S, Mazzeo D, Goult BT, Gingras AR, Pritchard CA, Critchley DR, Monkley SJ - Eur. J. Cell Biol. (2010)

Analysis of MEFs for talin structure function studies. (A) Western blots of Tln1fl/fl; CreER/+ MEF lysates 72 h after treatment with 100 nM 4-hydroxy tamoxifen (4OHT) or ethanol (EtOH) probed with talin 1 and 2 specific antibodies with vinculin as a loading control. (B) Numbers of GFP- or paxillin-containing FA in Tln1fl/fl; CreER/+ MEF 72 h after treatment with 100 nM 4OHT and either untransfected or transfected as shown. Results are expressed as mean ± s.e.m. (C) Different batches of primary MEFs differ in their phenotype before and after treatment with 4OHT. (Top panels) Brightfield images of two different Tln1fl/fl; CreER/+ MEF lines 96 h after treatment with either ethanol (EtOH) or 100 nM 4OHT. (Bottom panels) Western blot of lysates of same MEF lines 24 h or 96 h after treatment with either ethanol (−) or 100 nM 4OHT (+). Vinculin was used as a loading control.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2958305&req=5

d32e1464: Analysis of MEFs for talin structure function studies. (A) Western blots of Tln1fl/fl; CreER/+ MEF lysates 72 h after treatment with 100 nM 4-hydroxy tamoxifen (4OHT) or ethanol (EtOH) probed with talin 1 and 2 specific antibodies with vinculin as a loading control. (B) Numbers of GFP- or paxillin-containing FA in Tln1fl/fl; CreER/+ MEF 72 h after treatment with 100 nM 4OHT and either untransfected or transfected as shown. Results are expressed as mean ± s.e.m. (C) Different batches of primary MEFs differ in their phenotype before and after treatment with 4OHT. (Top panels) Brightfield images of two different Tln1fl/fl; CreER/+ MEF lines 96 h after treatment with either ethanol (EtOH) or 100 nM 4OHT. (Bottom panels) Western blot of lysates of same MEF lines 24 h or 96 h after treatment with either ethanol (−) or 100 nM 4OHT (+). Vinculin was used as a loading control.
Bottom Line: The talin rod contains several actin-binding sites (ABS), and mutations in the C-terminal ABS that reduced actin-binding impaired talin1 function, whereas those that increased binding resulted in more stable FAs.The results show that both the N-terminal integrin and C-terminal actin-binding functions of talin are essential to cell spreading and FA assembly.Finally, mutations that relieve talin auto-inhibition resulted in the rapid and excessive production of FA, highlighting the importance of talin regulation within the cell.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, University of Leicester, Leicester, UK.

Show MeSH
Related in: MedlinePlus