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. 2011 Sep 8;286(42):36378–36384. doi: 10.1074/jbc.M111.296483

FIGURE 3.

FIGURE 3.

Effect of MCAK depletion on the proliferation of paclitaxel-dependent cells. A, WT CHO cells and paclitaxel-dependent cell lines Tax 11-6 and HAβ3-3 were transfected with pHygro/U6 (Vector) or pHygro/U6-MCAK (shRNA) and plated for colony formation in hygromycin but without paclitaxel. B, extracts from untransfected HAβ3-3 and Tax 11-6 cells (lanes 1 and 3) and from the same cells that survived in the absence of paclitaxel following transfection with pHygro/U6-MCAK (lanes 2 and 4) were examined by Western blotting with antibodies to MCAK. An antibody to actin was included as a loading control. C, HAβ3-3 cells before transfection (−) and after survival following MCAK depletion (+) were also examined by Western blotting with antibodies to MCAK and the HA tag to ensure that loss of HAβ3-tubulin was not the reason they lost paclitaxel dependence.