Fig. 3

GRK6 palmitoylation and kinase activity determine the EMT progression and cellular migration activity in TNBC cells. A. The illustration for the difference of amino acid sequence (highlighted at pink) at the C-terminal region of palmitoylatable GRK6A (wild-type, wt) and non-palmitoylatable GRK6B (mutant, mt) variants. B–C. Western blot analyses for protein levels of membrane (Mem.)-associated/cytosolic (Cyt.)/total GRK6 and GAPDH in the parental (PT) HCC1937 cells without or with overexpression of vector control, GRK6A or GRK6B (B) and MDA-MB231 cells treated with GRK6 inhibitor GRK6-IN-2 at the indicated concentrations for 24 h (C). D–E. Western blot analyses (D) for GAPDH and EMT markers E-cadherin (E-Cad), N-cadherin (N-Cad), Fibronectin (FN), Vimentin (VIM) and Slug in the whole cell lysates, and Giemsa stain (E, left) for the migrated cells in the trans-well cultivation of HCC1937 cell variants. The migrated cell number from three independent experiments were presented as mean ± SEM in the histogram (E, right). F–G. Western blot analyses (F) for GAPDH and the indicated EMT markers in the whole cell lysates, and Giemsa stain (G, left) for the migrated cells in the trans-well cultivation of MDA-MB231 cells treated with GRK6-IN-2 at the indicated concentrations for 24 h. The migrated cell number from three independent experiments were presented as mean ± SEM in the histogram (G, right). In B, C, D and F, GAPDH was used as an internal control of protein loading. In E and G, the different alphabets indicate the statistical significance at p < 0.05