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. 2019 Jul 25;20(10):1337–1347. doi: 10.1080/15384047.2019.1638678

Figure 4.

Figure 4.

Depletion of kinesin-13s in A549 cells did not affect interphase array clearing in the presence of paclitaxel. (a) Depletion of kinesin-13’s, KIF2A and KIF2B, by siRNA knockdown was confirmed at 48 h post-transfection by immunoblotting. Gel samples were made concomitant with double thymidine synchronization at 24 and 48 h post-siRNA transfection. Actin served as a loading control. Attempts to detect KIF2C/MCAK by immunoblot were unsuccessful; however, KIF2A and KIF2B depletion appeared to be enhanced following triple transfection of KIF18A, KIF18B, and KIF2C/MCAK siRNA. (b) For DMSO-treated control A549 cells, microtubules were lost at the cell periphery independent of depletion of KIF2A and KIF2B. (c) In paclitaxel-treated cells, microtubule density in the central region of the cytoplasm also showed a similar reduction in early and late prophase for either control siRNA transfected cells or cells transfected with siRNA targeting the kinesin 13’s. For each of 3 independent experiments, at least 10 cells for each cell cycle stage of each experimental condition were measured. NT, Non-targeting; PTX, paclitaxel. **p < .01; ***p < .001; ****p < .0001.