Figure 4 – rAAV8 producing cells treated with nocodazole arrest in G2/M phase. Flow cytometry histograms triggered against propidium iodide signal for cells show the relative abundance of cells in either G1 (blue), S (orange) or G2/M (green) phases of the cell cycle in the absence (A ) or presence (B ) of 4 µM nocodazole addition at 0 HPT. Ratio of cells in G2/M phase to G1 phase at 72 HPT with 4 µM nocodazole added at 0, 4, 24, 48 HPT or untreated (Ctrl) (C ). Representative fluorescent composite images of rAAV-producing cells fixed 24 HPT show staining of the fibrillar component of the nucleolus with anti-fibrillarin (red) and nuclear DNA with DAPI (blue). Cells displaying disorganised nucleolar morphology are indicated by white arrows in non-treated cells (D ) and cells treated 4 hours post-transfection with 4 µM nocodazole (E ). Images taken using 100 x oil objective. Scale bar 10 µm. Data shown in (C ) are mean ± SEM for three biological replicates.