Journal: Neurophotonics

Article Title: Förster resonance energy transfer efficiency of the vinculin tension sensor in cultured primary cortical neuronal growth cones

doi: 10.1117/1.NPh.9.2.025002

Figure Lengend Snippet: (a) Box and whisker plot of FRET efficiency measurements in growth cones from neurons expressing VinTS and TSMod. *** denotes significance of p < 0.001 by Welch’s t -test. The cross (×) denotes the mean. The line denotes the median of the dataset, and the top and bottom of the box denote the 75th and 25th percentile, respectively. The black lines extending from the box (whiskers) corresponds to the maximum and minimum values, which are not considered outliers. Individual points above and below the whiskers are considered outliers with values larger than ± 1.5 × [the difference between the 75th and 25th percentile]. (b) The experiment in (a) was repeated to include VinTL as an additional control. *** denotes significance of p < 0.001 by ANOVA followed by Tukey’s multiple comparisons test. (c) Cumulative distribution functions for pixels from growth cones from neurons expressing VinTS and VinTL [data from (b) only].

Article Snippet: VinTS, VinTL, and TSMod plasmids were a gift from Martin Schwartz; VinculinTS (Addgene plasmid #26019; RRID:Addgene_26019) VinTL (Addgene plasmid #26020; RRID:Addgene_26020) and TSMod (Addgene plasmid #26021 RRID:Addgene_26021).

Techniques: Whisker Assay, Expressing

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

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 cDNA sequences for DN-KASH and VE-cadherin TSmod were subcloned into pShuttleCMV (Addgene plasmid 16405) and then recombined into pAdEasy-1 (Addgene plasmid 16400), following the AdEasy Adenoviral Vector System instructions (Agilent).

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