Figure 4
TACC1 interacts with RARα in the nucleus of F9 cells and is present in a nuclear chromatin-enriched fraction. (A) Cells from an F9 cell line stably expressing Flag-RARα were fractionated into cytoplasmic (C) and nuclear (N) fractions. 800 μg of total protein of each fraction were immunoprecipitated with an anti-Flag antibody. Immunoprecipitates were resolved by SDS-PAGE and blotted with anti-Flag or anti-TACC1antibodies, which reveal only the longest TACC1 proteins (A and K). Absence of cross contamination between cytosolic and nuclear fractions was verified using respectively anti-β-tubulin and anti-Histone H3 antibodies. The same amount of protein was loaded in each lane. (B) F9 cells were fractionated into cytosolic (lane 1), soluble nuclear (lane 2) and chromatin-enriched (lane 3) fractions. Cell-equivalent amounts of fractions were probed by immunoblotting with anti-TACC1, anti-RARα, and anti-Histone H3 antibodies.