Figure 5From: Target of rapamycin signaling regulates high mobility group protein association to chromatin, which functions to suppress necrotic cell deathReduced TORC1 signaling and the H3K37A mutation destabilizes Nhp10 binding to chromatin. (A) H3WT and H3K37A strains expressing Nhp10-6XHA were grown to log phase before mock treating or treating with 25 nM rapamycin for 30 minutes (upper panels) or 2 hours (lower panels). Samples were fractionated into soluble and chromatin-bound proteins and 30 μg of each was immunoblotted with the indicated antibody. Each set of immunoblots was exposed for the same length of time. Chromatin-bound Nhp10 and histone H3 were quantified with ImageJ software and expressed as a ratio of Nhp10/histone H3. The rapamycin samples were normalized to the mock controls, which were set to a value of 1. (B) As (A), except strains expressed Hmo1-6XHA. Only the 30-minute rapamycin treatment is shown, since Hmo1 protein levels decrease significantly by 2 hours post-rapamycin treatment. Rap, rapamycin; WT, wild-type.Back to article page