35S‐radiolabeled recombinant Myc‐Rab1a protein loaded with vehicle, GDP or GMP‐PNP was added to GST, GST‐C9orf72S, and GST‐C9orf72L immobilized on glutathione‐coated beads.
35S‐radiolabeled recombinant Myc‐Rab1a protein was visualized by phosphorimager (top panel). Coomassie‐stained GST, GST‐C9orf72S, and GST‐C9orf72L in the pull‐down samples are shown (bottom panel). The identity of the Coomassie protein bands was confirmed by mass spectrometry (
# indicates
E. coli DnaK chaperonin; * indicates
E. coli 60kD chaperonin;
Appendix Fig S3). Relative binding of Rab1a to C9orf72 was quantified from 3 independent experiments (mean ± SEM; one‐way ANOVA with Fisher's LSD test; ns, not significant; *
P ≤ 0.05; **
P ≤ 0.01; ***
P ≤ 0.001).