FIG. 3.

Hzf overexpression in fibroblasts causes the inhibition of cell proliferation. (A) Schematic representations of Hzf and mutant proteins. Full-length Hzf protein (WT) contains three C2H2-type zinc finger domains (black bars). (B) Colony formation assay. NIH 3T3 or p53-null fibroblasts were infected with control or Hzf retroviruses. After 3 days of infection, 10⁴ cells were seeded per plate, cultured for another 10 days, and stained with Giemsa. (C) NIH 3T3 cells were infected with indicated retroviruses and analyzed 3 days later for DNA content by FACS. (D) Cells were fixed and stained with Giemsa, and binucleate cells were counted at the indicated times postinfection. (E) After 3 days of infection, cells were stained using anti-γ-tubulin antibody, and the numbers of centrosomes per cell were counted. (F) Extranumerary centrosomes are present in cells 4 days after infection with an Hzf-expressing vector versus a vector control. (G) NIH 3T3 cells infected with the indicated retroviruses were pulse-labeled with BrdU for 30 min and stained with anti-BrdU antibody. (H) The same cells as shown in panel G were stained with annexin V and analyzed by FACS.