Fig 8. Kek-6 physically interacts with synaptic factors.

(A) Diagram of pull-down workflow and (B) coomassie-stained gel showing proteins from S2 cell or adult fly head lysate bound to Kek-6-Flag or mock control. Red boxes indicate fractions used for mass spectrometry. Hits that appeared in mock controls, or with only one peptide, were dis-regarded. (C) Selection of candidate interacting proteins, see S2 and S3 Tables. (D) Co-immunoprecipitations from co-transfected S2 cells: precipitating Kek-6-Flag brought down CaMKII-HA and VAP33A-HA. IP: immunoprecipitation, WB: western blot. (E) Western blots with anti-activated pCaMKII^T287, non-phosphorylated plain CaMKII and Tubulin, from adult fly heads, GMRGAL4 drives expression in retina. Graphs show pCaMKII^T287 levels normalized over non-phosphorylated CaMKII. Student paired t-tests. *p<0.05, **p<0.01, n = 5–9 replicates, see S1 Table.