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. 2015 Jun 16;108(12):2783–2793. doi: 10.1016/j.bpj.2015.05.010

Figure 6.

Figure 6

YAP signaling requires strain transfer to the nucleus through nesprin-1 giant. (A) Knockdown of nesprin-1 giant was performed using lentiviral delivery of miRNA, and knockdown was verified by dot blot for nesprin 1 after 1-MDa size filtration. (B) Quantification of the nuclear deformation index with knockdown of nesprin-1 giant compared to control cells (miRneg). Positive index indicates deformations larger than control, negative index indicates deformations less than control (mean ± SE, n = 146–149 cells/group). With less nesprin-1 giant to connect the nucleus to the cytoskeleton, smaller deformations occur. (C) Images of YAP (bottom) and DAPI (top) stained nuclei on aligned scaffolds with or without 30 min of dynamic tensile loading (DL, 3% strain; 1 Hz, scale = 25 μm) and with or without nestrpin-1 giant knockdown. (D) Quantification of the nuclear/cytoplasmic YAP for control (noted as “C”) or dynamically loaded (DL) cells. (Mean ± SE, n = 33–41 cells/group; # p < 0.05, ∗p < 0.05, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.) To see this figure in color, go online.