EB1 targets KIF17 to MTs. (A) Immunostaining of endogenous KIF17 and transfected myc-EB1-FL or myc-EB1-FLEE in Caco2 cells. Color overlays show boxed regions at higher magnification. (B) Line-scan analysis of the number and fluorescence intensity of KIF17 puncta along the distal 10 µM of individual MTs in cells expressing myc-EB1-FL or myc-EB1-FLEE. Data derived from three experiments in which 487 and 339 puncta along 42 MTs were analyzed in cells expressing myc-EB1 wild-type (WT) or mutant (EE) proteins. (C) Immunostaining of KIF17 and tyrosinated MTs in EB1-depleted cells. Color overlay shows an enlargement of the boxed regions. Arrows, MT ends decorated by KIF17; arrowheads, MT ends not decorated by KIF17. Western blot shows EB1 and tubulin levels in siNC or siEB1 transfected cells. Graph shows quantification of MT ends decorated by KIF17 in siNC or siEB1 cells (n = 3,058 and n = 3,882, respectively, compiled from three experiments). Error bars indicate SEM. (D) Immunostaining of tyrosinated MTs and EB1 in control and KIF17-depleted cells. Western blot shows KIF17 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) levels in Caco2 cells infected with lentiviral control (shNC) or KIF17 shRNAs (shKIF17#1 and shKIF17#2 target two unique sequences).
