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Fig. 3 Alcohol suppresses de novo rRNA synthesis and content in O9-1 cells and primary chick CNCs. (A) Alcohol exposure at the EC50 for O9-1 cells (80mM) and chick primary CNCs (52mM) reveals fewer or no EU-positive nuclei (red) in alcohol-exposed nuclei (blue) as compared with controls. Nuclei visualized using DAPI; bar indicates 50 μm. (B) Dose-response shows a significant decline in de novo rRNA synthesis, quantified using qPCR for ITS-1, at alcohol exposures at or above 40mM for O9-1 cells. (C) At 30min following exposure to 80mM alcohol there is a significant decline in ITS-1 content in O9-1 cells. (D) At 30min following exposure to 52mM alcohol there is a significant decline in ITS-1 content in chick primary CNCs. Values are mean ± SD for three independent replicates. Data analyzed using two-tailed Student’s t-test (A) or one-way analysis of variance (B-D).