Figure 6
Depletion or overexpression of CEP20 changes the status of microtubule polymerization in A549 cells. (A and B) Lysates from A549 cells transfected with the indicated plasmids were subjected to ultracentrifugation. The supernatant (S) and pellet (P) fractions were then processed for Western blotting with anti-α-tubulin and CEP20 antibodies. Uncropped immunoblots were shown in Supplementary Fig. S8D. The intensities of the bands were quantified by ImageJ. (C and D) A549 cells transfected with GFP or GFP-CEP20 were processed for immunoflourescence assay with anti-α-tubulin antibody. DNA was stained with DAPI (blue). Bar, 10 μm. The average intensity of microtubules in cells (n = 40) transfected with GFP or GFP-CEP20 was determined by ImageJ. Quantitative data of microtubule intensities are presented as mean ± SD. (E and F) Lysates from A549 cells treated with nocodazole and CEP20 depletion were subjected to ultracentrifugation. The supernatant (S) and pellet (P) fractions were then processed for Western blotting with anti-α-tubulin. Uncropped immunoblots were shown in Supplementary Fig. S8E and F. Quantitative data of polymerized tubulin are presented as mean ± SD. *P < 0.05 and **P < 0.01, student’s t-test.
