Figure One - Lysates of A549 WT cells and A549 Rab8A KO cells, 20 µg of protein was loaded per lane. The antibodies were used in 5% milk in TBST at 1 ug/ml. The membranes were incubated with the antibodies overnight in 4°C. The secondary sheep antibody conjugated with HRP was used in 5% milk in TBST for 1 hour at RT. Figure Two - For IP with S969D 200 µg of Ab was coupled with 1000 µl of crude beads and then 30 µl of beads used per 1 mg of protein. IP was performed for 3 h at 4°C. Lysates of A549 WT and A549 Rab8A KO were used.