Nmiia

The primary antibodies used were: a) rabbit polyclonal – NMIIA (Sigma), phospho-NMIIA (Ser1943) (Cell Signaling Technology), giantin, Man-II, and Rab6a (Abcam); b) mouse monoclonal - Rab6a (Santa Cruz Biotechnology), β-actin (Sigma), Sar1a, giantin (Abcam); c) mouse polyclonal - ST3Gal1 (Abnova). The secondary antibodies (Jackson ImmunoResearch) were: a) HRP-conjugated donkey anti-rabbit and donkey anti-mouse for Western-blotting; b) donkey anti-mouse Alexa Fluor 488 and anti-rabbit Alexa Fluor 594 for immunofluorescence. Blebbistatin (Sigma) and latrunculin A (Tocris Bioscience) were dissolved in dimethyl sulfoxide (DMSO) immediately before use. Cells treated with a corresponding concentration of DMSO served as controls. The regular working concentrations were: Blebbistatin −25 μM for up to 24 h; latrunculin A −0.2 μM for up to 24 h. Active human Rab6a full length protein was obtained from Abcam.

(−)‐Blebbistatin, primaquine and phenylephrine were purchased from Sigma. siRNA transfection reagents were purchased from Horizon Discovery. DPBA was purchased from Santa Cruz Biotechnology. The following primary antibodies were used: NMIIA (rabbit, 1:200), NMIIB (rabbit, 1:200), GAPDH (rabbit, 1:20 000) and α‐smooth muscle actin (mouse 1:10 000) from Sigma; β‐Cytoplasmic actin (mouse 1:500) and γ‐Cytoplasmic actin (mouse 1:500), a gift from Christine Chaponnier; phospho‐FAK Y925 (rabbit, 1:200) and phospho‐Paxillin Y118 (rabbit, 1:250) from Cell Signaling Technologies; FAK monoclonal antibody (mouse, 1:250) from Thermo Fisher Scientific; anti‐Paxillin (mouse, 1:500) from BD biosciences; anti‐actin antibody (rabbit, 1:500) from Cytoskeleton. For immunofluorescence experiments, goat anti‐mouse and goat anti‐rabbit Alexa Fluor 488 and Alexa Fluor 568 (1:1000, Invitrogen) were used as secondary antibodies. For Western blotting experiments, IRDye 680 RD and IRDye 800CW labelled goat anti‐rabbit and goat anti‐mouse (LI‐COR, 1:10 000) were used as secondary antibodies.