FIGURE 2.

Effects of ROCK‐is on physical stiffness of the 3T3‐L1 3D spheroids. At Day 12, the 3D human orbital fibroblast (HOF) spheroids of preadipocytes (DIF−) and their adipogenic differentiation (DIF+) without or with 1 or 10 µM ripasudil (Rip), Y27632, or KD025 were subjected to a physical solidity analysis using a microsqueezer. The force required to induce deformation until half diameter was reached (μN/μm force/displacement) were measured and the data are potted in panel A. Among the different ROCK‐is forms, to compare their effects toward HOFs adipogenesis, the physical stiffness of 10 µM of each ROCK‐is were replotted in panel B. All experiments were performed using freshly prepared 12–20 spheroids. *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001 (ANOVA followed by Tukey's multiple comparison test). ROCK, Rho‐associated coiled‐coil containing protein kinase