Fig. 9
Rab14 interacts with ADAM10 and, upon SC79 treatment, moves to the cell surface. (A) Rab14 co-immunoprecipitates with ADAM10. HAECs were treated with DMSO or SC79 (10 µM) for 20 min before being lysed with NP-40 buffer. Lysates were incubated with rabbit anti-ADAM10 antibody or rabbit IgG as a control overnight at 4 °C, followed by incubation with Protein A agarose. The input and immunoprecipitation fractions were analyzed using anti-ADAM10 and anti-Rab14 antibodies. (n = 4, *p < 0.05 vs. cells treated with DMSO and immunoprecipitated with ADAM10 antibody, ns: not significant). (B) SC79 translocates ADAM10 and Rab14 from intracellular compartments to the cell surface. HAECs were treated with DMSO or SC79 (10 µM) for 20 min. The cell surface proteins were biotin-labeled and isolated with avidin-coated agarose beads. Whole cell lysates, biotin-bound cell surface proteins, and biotin-unbound intracellular proteins were examined by Western blotting using anti-ADAM10, Anti-Rab14, and anti-actin antibodies. (n = 5, *p < 0.05)
