Figure 5.

Alpha-synuclein A53T mutation affects the strength of the interaction of α-syn with RAN. (A) Western-blot analysis of homogenates from A53T and A53T-corrected hiPSC-derived MD NPC lines. Anti-syn immunoprecipitates were analyzed by western blot using anti-RAN antibody in unbound and bound fractions. The immunostaining with β-actin antibody was used as the reference for the unbound fractions. The results were calculated as the ratio between bound and unbound fractions for RAN protein and normalized to β-actin expression in the same lane. RAN bands visualized at 25 kDa, β-actin (42 kDa) was used as internal control for normalization in the unbound fraction. (B) Ratios of the bound compared with the unbound levels of RAN showed stronger interaction for the A53T mutation compared with the isogenic mutation corrected control (^*P < 0.05, Student’s t-test).