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. 2007 Jan 15;176(2):183–195. doi: 10.1083/jcb.200610099

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Copyright © 2007, The Rockefeller University Press

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Figure 7. — Depleting APC affects the mitotic checkpoint and apoptosis and leads to tetra- and polyploidy in HCT116 cells. (A) HCT116 cells transfected with control (first lane) or APC-targeting (second lane) siRNA for 36 h were lysed, separated by PAGE, and blotted for APC and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; loading control). (B) APC inhibition results in mitotic spindle checkpoint defects in HCT116 cells. HCT116 cells were transfected with control (purple) or APC-targeting (pink) siRNA for 36 h and were either arrested with 10 ng/ml taxol or 100 ng/ml nocodazole for 12 h or were left untreated. Cells were harvested, fixed, and stained with phosphohistone H3 antibody to detect mitotic cells. (C) The G1 tetraploid population is increased in APC-deficient HCT116. HCT116 cells transfected and treated as in B were harvested and stained for cyclin B1 and DNA content as in Fig. 3. (D) The polyploid population is increased in APC-deficient HCT116 cells. The graph represents the polyploid population in control (purple) and APC-deficient HCT116 cells treated and stained as in C. (E) Apoptosis is decreased in APC-deficient HCT116 cells. The sub-G1 fractions in control (purple) and APC-deficient HCT116 cells treated and stained as in C are shown. (B–E) Values are given as the mean ± SD (error bars).