Figure 3.

Depletion of PAF49 does not cause cell cycle arrest in a specific phase.A, FACS analysis of cells at the indicated times post-treatment of IAA. B, quantification of the distribution of cells in G₁, S, or G₂ following IAA treatment. Four independent repeats of the analyses presented in (A) were each carried out in duplicate. The data were analyzed by two-way ANOVA with Tukey’s multiple comparisons test. C, ratio of cells in S and G₁ was calculated for the data presented in (B). Significance was determined by a one-way ANOVA with Tukey’s multiple comparisons test. D, Cells containing AID-PAF49 were treated with 1 mM IAA or vehicle for up to 6 days. On days 0, 2, 4, and 6, cells were harvested and lysed in HEPES lysis buffer. The expression of p53 was determined by Western blot analysis with anti-p53 antibody. E, three independent repeats of the Western blot data presented in (D) were performed. The levels of p53 were corrected for the signal of the housekeeping protein β-actin. The data were analyzed by one-way ANOVA with Dunnett’s multiple comparisons test. The error bars represent mean ± SD; ∗ = 0.05 to 0.01, ∗∗ <0.01, ∗∗∗ <0.001, ∗∗∗∗ <0.0001. F, cells containing AID-PAF53 were treated with 500 μM IAA for 3 h and 20 h. Reduction of PAF53 shows reorganization of nucleolar structures into caps along the periphery, where UBF and fibrillarin colocalize and RPA194 disperses into the nucleoplasm. Scale bar = 5 μm.