(A) Representative images of YAP/TAZ and Lamin-A/C immunostaining of MSCs cultured in 9 kPa MMP-degradable and non-degradable NorHA hydrogels for 5d prior to fixation and staining, along with the corresponding quantifications of (B) nuclear shape factor and (C) nuclear volume (n=13–23 nuclei/group, * = p <0.05 by Student’s t-test). SB = 5 μm. (D) YAP/TAZ nuclear-to-cytoplasmic ratios of MSCs from A-C (n=13–22 cells, Box plot represents median and 25th/75th quartiles, **** indicates p<0.0001 by Mann-Whitney U-test). (E-G) Quantifications of 3D nuclear metrics including nuclear shape factor (E), nuclear volume (F), and nuclear wrinkling index (G) as compared to YAP/TAZ nuclear-to-cytoplasmic ratios of MSCs from A-C (n=13–22 cells, ** indicates p<0.01 from Pearson’s correlation test, r and p-values shown for MMP-degradable hydrogel fit).