Fig. 4

Rho activation by MLN triggers MAT and tumoroid spreading in PC3 cells. A Activation of Rho isoforms by 100 nM MLN in PC3 cells. The active GTP-bound isoforms are hash-tagged. B Depletion of Rho isoforms in PC3 cells changes cell morphology and affects the cell response to 100 nM MLN. Scale bar = 25 µm. C Quantitative analysis of the effect of Rho isoform knockdown shown in (B). The cell area and length-to-width ratio (LWR) were measured by automated microscopy. Statistical significance: ***-p < 0.001. D Effect of the depletion of Rho isoforms on PC3 tumoroid growth. Scale bar = 200 µm. E Effect of 10 µM Y27632, 20 µg/ml C3E, and 3 µM CCG1423 on PC3 tumoroid growth with or without (control) 100 nM MLN. Scale bar = 200 µm. F Localization of RhoC in control and 100 nM MLN-treated PC3 cells. Scale bar = 25 µm and 10 µm (zooms). In all experiments with MLNs, DMSO was used as a vehicle control