Fig 2: Effect of KIF11 or KIF15 depletion on growth of U-R or H-R cells.(A) Experimental procedure. (B) Evaluation of knockdown efficiency by immunoblotting. (C) KIF11 knockdown heavily compromised the growth of the U-R cells, while the growth of H-R cells was affected only when both KIF11 and KIF15 were depleted, suggesting their mutually compensatory relationship. Cell viability was shown relative to that for the cells treated with control siRNA.

Fig 3: Ectopic expression of KIF11S1017fs leads to moderate STLC resistance in U-2OS cells.(A) Immunoblot of the U-2OS cells expressing the KIF11 transgenes. (B) STLC sensitivity of the U-2OS cells expressing the KIF11 transgenes. Relative cell viability for a given STLC concentration is shown as mean ± SD (n = 5). Lentivirus-mediated transduction of KIF11S1017fs rendered U-2OS moderately resistant to STLC, to the same degree as with the KIF11G268V allele [8]. Overexpression of the wild-type allele failed to confer any notable resistance. (C) STLC sensitivity of the HeLa cells expressing the KIF11 transgenes.

Fig 4: U-R cells express kinesin-5 with higher mobility.(A) The amount of KIF11 and KIF15 protein in the U-R and H-R cells was evaluated by immunoblotting. KIF15 was moderately increased in H-R cells relative to the parental HOS cells. Intriguingly, U-R cells expressed a KIF11 variant that migrated slightly faster on SDS-PAGE, in addition to normal KIF11. (B) mRNAs for KIF11 and KIF15 were quantified using qPCR and the results from five independent experiments were shown as mean ± SD. Two-tailed t-tests were used for statistical evaluation. KIF15 mRNA in the H-R cells was significantly increased in comparison with the parental HOS cells, whereas no quantitative change in the U-R clone 9 was observed. For KIF11 mRNA, neither the U-R cells nor the H-R cells showed any change in the expression level.

Fig 5: DR6 functions through its association with KIF11 and TRAF4. (A) Overexpression of KIF11 or TRAF4 reversed the DR6 knockdown‐mediated reduction in SKOV3 cell migration. Scale bar = 200μm. (B) Knocking down of KIF11 or TRAF4 reversed the DR6 overexpression‐mediated increase in SKOV3 cell migration. Scale bar = 200μm. (C) Western blot analysis of indicated protein expression in SKOV3 cells. Data are presented as mean ± SD (n = 8). **P < 0.01 (Student's t test).