Inhibition of PKCα results in AMT in amoeboid cells, PKC activation results in MAT in mesenchymal cells. Cell morphology analysis of A375m2 (A), K2 (B) and MDA-MB-231 (C) strains upon PKC activator PMA or PKCα/β inhibitor Gö6976 treatment. Cells were incubated with PMA (162nM) or Gö6976 (1 μM) for 48 hours in 3D collagen and then analyzed using NIS Elements software (Nikon). Cells were classified according to the elongation index as rounded, intermediate or elongated (see Methods section). Statistical significance (marked by asterisks) was evaluated using Pearson’s Chi-squared test. Representative results are shown out of minimum three independent experiments.