Fig 3.

LANA preferentially binds to the nonmethylated N-terminal H3 tail. Peptide pulldown analyses were performed by incubation of 1 μg of biotinylated peptides with purified LANA, luciferase, or EHMT1. Biotinylated peptides were precipitated with a streptavidin bead mixture and washed extensively with binding buffer containing 500 mM NaCl. Interaction with the histone N-terminal tail was probed by immunoblotting with anti-Flag antibody. The EHMT1 protein was used in the peptide pulldown assay as a positive control for H3K9me1/2 binding, while luciferase protein served as a negative control. LANA but not luciferase interacts with the histone H3 N-terminal tail in a posttranslational-modification-specific manner. The band intensities are shown as a percentage of the input.