FIGURE 5.
Super shift analysis with anti-PARP-1 antibody for reconfirmation of PARP-1 binding to the T−172 probe. Each sample, consisting of 1.0 μg of nuclear protein extracted from SH-SY5Y (lanes 1–4) or HEK293T (lanes 5–8), was loaded in EMSA. Anti-PARP-1 antibody was prepared after the dialysis and the concentration because commercially available antibody was too thin. Salt concentration of anti-PARP-1 antibody, concentrated dialyzed buffer, and anti-SP1 antibody was equal in respective sample. Lanes 1 and 5, nuclear extract without any antibody or dialyzed buffer; lane 2 and 6, nuclear extract with 2.0 μl of dialyzed buffer used for dialysis of antibody solution; lanes 3 and 7, nuclear extract with 2.0 μl of anti-PARP-1 antibody; lanes 4 and 8, nuclear extract with 2.0 μl of anti-SP1 antibody as a negative control (CON). These results were performed of at least three independent experiments.