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Disruption of the LINC complex affects focal adhesion dynamics. (A) DN-KASH affects focal adhesion morphology. Cells expressing AV-mCherry or AV-DN-KASH-mCherry were immunostained for DAPI, <t>vinculin,</t> or vimentin (green) and actin (gray). (B) DN-KASH expression affects the vimentin cytoskeleton. Cells expressing AV-mCherry or DN-KASH-mCherry were immunostained for DAPI, vimentin (green), and actin (gray). (C) Cells were cotransfected with <t>vinculin-TSmod</t> and AV-mCherry or DN-KASH-mCherry. Force exerted across vinculin is reduced upon LINC disruption in HUVEC. Coinfected cells with adenovirus-mCherry or DN-KASH-mCherry with vinculin tension-sensor (TSmod). Mann–Whitney test (* p < 0.05), N = 3. (D) Altered phosphorylation states of focal adhesion proteins with DN-KASH expression. HUVEC-expressing AV-DN-KASH or AV-mCherry control were lysed for 36–48 h and blotted for pY1065 vinculin, pS425 Talin, and pY397 FAK ( N = 3). Mann–Whitney test (** p < 0.01).
Vinculin Tsmod, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Disruption of the LINC complex affects focal adhesion dynamics. (A) DN-KASH affects focal adhesion morphology. Cells expressing AV-mCherry or AV-DN-KASH-mCherry were immunostained for DAPI, vinculin, or vimentin (green) and actin (gray). (B) DN-KASH expression affects the vimentin cytoskeleton. Cells expressing AV-mCherry or DN-KASH-mCherry were immunostained for DAPI, vimentin (green), and actin (gray). (C) Cells were cotransfected with vinculin-TSmod and AV-mCherry or DN-KASH-mCherry. Force exerted across vinculin is reduced upon LINC disruption in HUVEC. Coinfected cells with adenovirus-mCherry or DN-KASH-mCherry with vinculin tension-sensor (TSmod). Mann–Whitney test (* p < 0.05), N = 3. (D) Altered phosphorylation states of focal adhesion proteins with DN-KASH expression. HUVEC-expressing AV-DN-KASH or AV-mCherry control were lysed for 36–48 h and blotted for pY1065 vinculin, pS425 Talin, and pY397 FAK ( N = 3). Mann–Whitney test (** p < 0.01).

Journal: Molecular Biology of the Cell

Article Title: The LINC complex is required for endothelial cell adhesion and adaptation to shear stress and cyclic stretch

doi: 10.1091/mbc.E20-11-0698

Figure Lengend Snippet: Disruption of the LINC complex affects focal adhesion dynamics. (A) DN-KASH affects focal adhesion morphology. Cells expressing AV-mCherry or AV-DN-KASH-mCherry were immunostained for DAPI, vinculin, or vimentin (green) and actin (gray). (B) DN-KASH expression affects the vimentin cytoskeleton. Cells expressing AV-mCherry or DN-KASH-mCherry were immunostained for DAPI, vimentin (green), and actin (gray). (C) Cells were cotransfected with vinculin-TSmod and AV-mCherry or DN-KASH-mCherry. Force exerted across vinculin is reduced upon LINC disruption in HUVEC. Coinfected cells with adenovirus-mCherry or DN-KASH-mCherry with vinculin tension-sensor (TSmod). Mann–Whitney test (* p < 0.05), N = 3. (D) Altered phosphorylation states of focal adhesion proteins with DN-KASH expression. HUVEC-expressing AV-DN-KASH or AV-mCherry control were lysed for 36–48 h and blotted for pY1065 vinculin, pS425 Talin, and pY397 FAK ( N = 3). Mann–Whitney test (** p < 0.01).

Article Snippet: The design of VE-cadherin TSmod ( Conway et al. , 2013 ) (Addgene plasmid 45848) and vinculin TSmod ( Grashoff et al. , 2010 ) (Addgene plasmid 26019) were previously described.

Techniques: Disruption, Expressing, MANN-WHITNEY, Phospho-proteomics, Control