Fig. 7.

RhoA-ROCK inhibition increases mineralization and EphrinB2 inhibits autophagy through a RhoA-ROCK-dependent mechanism. a–d Mineralization by Kusa 4b10 cells is enhanced when RhoA-ROCK is inhibited with either H1152 or Y27632. Kusa 4b10 osteocytes were cultured under mineralizing conditions and treated with H1152 (a, b) or Y27632 (c, d) at the doses indicated for the duration of the culture. Alizarin Red staining and quantitation is shown. Data are mean ± SD of three replicates from a representative of three independent experiments; **p < 0.01 vs. untreated (0) at same day of differentiation by two-way analysis of variance (ANOVA). e Autophagy is suppressed by EphrinB2 in a RhoA-ROCK-dependent manner. LC3 punctae formation in Ocy454 cells, either untreated or treated with bafilomycin A1 (Baf) (50 nM), in the presence and absence of clustered EphrinB2-Fc (1 μg/mL) and H1152 (50 μM) for 1 h. Punctae number per cell from two independent experiments, each performed in duplicate, with 95% confidence interval (CI); ***p < 0.0001; **p < 0.001 for comparisons shown, by one-way ANOVA with correction for multiple comparisons