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Fig. 6. Inhibition of HBP expression results in delay at G₁/S phase boundary and interferes with S phase progression. HeLa cells were transfected with either pGEM-U6pHP1 or pGEM-U6pT and arrested by HU treatment as described in Materials and Methods. Samples were prepared for FACS analysis by staining with propidium iodide at the indicated times after release from HU. (A) FACS profiles. Time points shown are hours after release from HU-treatment. Zero-hour samples were taken immediately before the release from HU block. The boundaries that define cells with G₀/G₁, S- and G₂/M-phase DNA content were determined as described in Fig. 4A. (B) Cell cycle distribution. Shown is the percentage of cells with G₀/G₁, S- and G₂/M-phase DNA content. (C) Analysis of protein samples by western blotting. Membranes were probed with anti-cyclin A, anti-HBP and anti-tubulin antibodies except for the 0-hour time point when samples from the same protein preparation were analysed on different gels. Note that therefore the 3, 8 and 16 hours anti-tubulin panels are identical. Results are representative of three independent experiments.

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