Fig. 3: KIFC1 is a novel TACC3 binding protein that is involved in TACC3-mediated CC.

A, B Correlation of KIFC1 mRNA expression with TACC3 (A) and CC score (B) in breast cancer patients with high CA20 expression in METABRIC dataset. C–E Distant relapse-free survival (DRFS) analyses in breast cancer patients with high CA20 expression based on TACC3 or KIFC1 or their combination in GSE22219 dataset. F IF staining of TACC3 (green)/KIFC1 or γ-tubulin (red) in JIMT-1 cells, showing colocalization at the centrosomes. Scale bar = 10 µm. G APEX2 proximity ligation assay to show binding of TACC3 to its interactors. H Western blotting of KIFC1 and the known interactor, clathrin upon TACC3 pulldown in APEX2-TACC3 overexpressing mitotic JIMT-1 cells. I IP of endogenous TACC3 and its interactors, KIFC1 and clathrin in JIMT-1 cells synchronized at mitosis and treated with BO-264 for 4 h (5 µM). J IF staining of α- (green) and γ- (red) tubulin in JIMT-1 cells transfected with siTACC3 or siKIFC1 (100 nM) for 48 h. Scale bar = 10 µm. K IF staining of α- (green) and γ- (red) tubulin in MCF-7 cells upon TACC3 + KIFC1 overexpression followed by CA induction with cytochalasin D. Scale bar = 10 µm. L Percent growth inhibition in MCF-7 cells upon TACC3/KIFC1 overexpression followed by CA induction with cytochalasin D. M A scheme of different truncated vectors of TACC3. N Quantification of the band intensities from O. O IP of KIFC1 and immunoblotting (IB) with GFP antibody in mitotic HEK293T cells transfected with GFP-labelled vectors of different regions of TACC3. 1-838: full length, 1-593: N-terminus, 594-838; C-terminus. P DARTS assay showing binding of BO-264 to the C-terminal TACC domain of TACC3.