Fig. 4: RGD/integrin and HAVDI/N-cadherin ligations regulate the adhesive state and the cytoskeletal organization of hMSCs.

a Left: Representative paxillin images in hMSCs on “OFF”, “ON” and “Dual ON” substrates for 1 d. Zoomed regions correspond to the rectangles marked in white in the main images. Right: Corresponding quantification of paxillin adhesion length (from left to right n = 146, 173, 154 adhesions in 41, 47, 45 cells respectively). b Left: Representative heat maps of traction stress in hMSCs on “OFF”, “ON” and “Dual ON” substrates for 1 d. Right: Corresponding quantification of average traction stress per cell (from left to right n = 37, 44, 47 cells). c Left: Representative F-actin images in hMSCs on “OFF”, “ON” and “Dual ON” substrates for 1 d (top). Zoomed regions display details of F-actin organization in the apical region of the nucleus (middle). The cross-sectional side view of lamin A/C staining was captured along the XZ-plane crossing the center of the nucleus (bottom). Right: Corresponding quantification of the percentages of hMSCs having the actin cap (top) (n = 3 experiments per group). Corresponding quantification of nuclear height (bottom) (from left to right n = 43, 55, 48 cells). d Schematic of a possible pathway for YAP nuclear translocation in response to the RGD/integrin and HAVDI/N-cadherin ligations. RGD on “ON” substrates induced integrin clustering and promoted formation of focal adhesions (FA) and actin cap. Contractile force from actin cap compressed and flattened the nucleus, and may have thereby enabled nuclear localization of YAP. HAVDI on “Dual ON” substrates reduced integrin clustering and FA formation, possibly attenuating nuclear compression and nuclear localization of YAP. Data are presented as mean ± s.e.m., and p values were obtained using one-way ANOVA followed by Tukey’s post hoc test (a–c). Scale bars: 20 µm (a), 30 µm (b), 30 µm for F-actin images and 5 µm for lamin A/C images (c). Source data are provided as a Source Data file.