Western Blot Analysis of Protein Extracts

Total protein extracts were prepared as described (35 (link)). Proteins were separated on 10% tris-glycine SDS-PAGE gels, transferred to 0.45 µm nitrocellulose membranes (Bio-Rad), and probed overnight at 4°C with mouse anti-HA (1:5,000; Sigma-Aldrich, 12CA5), rabbit anti-V5 (1:5,000; Invitrogen, MA5-15253), mouse anti-c-Myc (1:2,000–1:5,000; Sigma-Aldrich, 9E10), or rabbit anti-PSTAIR (1:5,000; Millipore-Sigma, 06-923). The rabbit polyclonal anti-H3 antibody was generated by Pocono Rabbit Farm & Laboratory and used at 1:100,000 dilution. Secondary anti-mouse and anti-rabbit antibodies conjugated to horseradish peroxidase were from Jackson ImmunoResearch (115-035-003 or 111-035-003) and used at 1:10,000 dilution for 60 min at 4°C. Immunoblots were developed with Clarity Western ECL Substrate (Bio-Rad, 170-5060) and imaged on a ChemiDoc MP multimode imager (Bio-Rad).