Fig. 1

Schematic diagram and western blot with quantitation. A RSV Gag constructs. B Subcellular fractionations were performed to separate the cytoplasm and nucleoplasm from the chromatin fractions, which were further separated using differential NaCl concentrations. The 150 mM chromatin fraction (Chr 150 mM) contains proteins associated with open chromatin (euchromatin). The 500 mM chromatin fraction (Chr 500 mM) contains proteins that are associated with condensed chromatin (heterochromatin). Wild type RSV Gag and Gag.L219A were detected in all of the fractions at different ratios. Gag.ΔNC was primarily detected in the cytoplasm with very little in the chromatin fractions. C Band densities were determined for each Gag construct for each fraction and are displayed as mean ± standard error of the mean (SEM) for three biological replicates. To assess fraction purity, cellular proteins were detected using antibodies against calnexin (cytoplasm), Med4 (nucleoplasm and euchromatin), and Histone H2B (euchromatin and heterochromatin). The position of molecular weight markers, in kilodaltons, are indicated on the right