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. 2019 Mar 4;13(3):2913–2926. doi: 10.1021/acsnano.8b06998

Figure 3.

Figure 3

Nanoneedles reduce YAP activity and lessen the correlation between YAP activation and cell spreading. (A) Confocal microscopy shows nuclear YAP protein localization on flat substrates and cytosolic localization on nN (green: YAP). Scale bars = 50 μm. (B) Image analysis quantification of YAP localization shows significant reduction in the nuclear to cytoplasmic ratio of YAP on nN (minimum/maximum; N = 4). (C) qPCR analysis indicates reduced expression of the YAP target genes ankyrin repeat domain 1 (ANKRD1) and connective tissue growth factor (CTGF) (N = 4, mean ± SD). (D) Cell spread area and YAP nuclear localization correlate tightly on flat substrates, but correlation is weakened on nN (N = 3). (E) YAP localization following cell treatment with either the actin depolymerizing agent, LatB, or a small molecule to stimulate actin bundling, LPA. LatB treatment on flat substrates reduces nuclear YAP localization to levels comparable to untreated cells on nN. LatB treatment of cells on nN yields a small reduction in nuclear localization. LPA treatment on flat substrates did not affect YAP localization for HUVECs and marginally decreased this metric for hMSCs. On nN substrates, LPA had little effect on YAP localization. (minimum/maximum; N = 3); *p < 0.05, **p < 0.01, ***p < 0.001 between groups as indicated by the lines.