Figure 4.

Effects of ROCK-i and/or BRI on adipogenic differentiation (DIF+) in 3D 3T3-L1 spheroids. The 3T3-L1 cells were 3D-cultured under several conditions: preadipocytes of 3T3-L1 cells (DIF−) and their adipogenic differentiation (DIF+) with or without 10 µM ripasudil (Rip) or KD025, and/or BRI. These were stained with BODIPY (A, scale bar: 100 μm), and their staining intensities were plotted (B). The mRNA expressions of DIF+-related genes, including Pparγ and Fabp4, under the above conditions were plotted in (C) 1–3. All experiments were performed in triplicate using fresh preparations, each consisting of five samples. Data are presented as the arithmetic mean ± standard error of the mean (SEM). * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001, and other pairs were not statistically significant (ANOVA followed by a Tukey’s multiple comparison test).