Fig. S7.

RABIF stabilizes Rab10 protein in E. coli. The RABIF gene was cloned into the pET28a-based SUMO vector (1), while Rab10 was cloned into the pGEX-4T3 vector. GST-Rab10 was expressed in BL21 E. coli cells either alone or together with RABIF. After the addition of 1 mM isopropyl β-d-1-thiogalactopyranoside (IPTG), the cells were incubated at 37 °C for 3.5 h. After cell lysis and centrifugation, Rab10 proteins in the supernatant were precipitated using glutathione beads. Rab10 proteins from the supernatant and whole cells were probed by immunoblotting using anti-Rab10 antibodies.