Figure 6.

ROCK, Cdc42, and Rac1 mediate Zc3h10 activity on mitochondrial networks. (A) Volcano plot of differentially expressed pre-mRNAs in ShZc3h10 and Scramble cells (36 h after differentiation induction). Red dots indicate labeled downregulated genes in ShZc3h10 preadipocytes regulating F-actin organization by Rho GTPases activity. Scramble: n = 2; ShZc3h10: n = 3; false discovery rate (FDR) < 0.05. (B) Schematic representation of the RhoA/ROCK pathway and the mechanism of action of Y27632. PKA inhibits RhoA, leading to reduced contractility of F-actin stress fibers and, ultimately, induces adipogenesis. (C) Schematic representation of the Rac1/Cdc42 pathway and the mechanisms of action of ML141 and NSC23766. PKA induces both Rac1 and Cdc42 activities, leading to peripheral F-actin remodeling in preadipocytes that favors adipogenesis. (D, F, and H) Representative confocal images of mitochondria in preadipocytes (36 h) that were exposed to 10 µM Y27632 (D), 10 µM ML141 (F), and 50 µM NSC23766 (H), and stained with Mitotracker Deep Red FM probe. Scale bar: 10 µm. (E, G, and I) Mitochondrial network analysis of the cells treated with 10 µM Y27632 (E), 10 µM ML141 (G), and 50 µM NSC23766 (I). n = 8 cells; one-way ANOVA followed by Tukey’s post-hoc test; *P < 0.05, **P < 0.01, ***P < 0.001.