GATA-1 and p53 interact in vitro in MEL cell extract. (A) A total of 320 μg of MEL nuclear extract was incubated with the following: lane 1, 4 μL of anti-GATA-1 N6 and 1 μL of anti-rat IgG2a; lane 2, 4 μL of anti-p53 (PAB 240) and 1 μL of anti-mouse IgG; lane 3, 5 μL of anti-rat IgG2 for 1.5 hours at 4°C with rotation and 40 μL of protein G plus agarose was added with 4 additional hours of rotation at 4°C; lane 4, MEL nuclear extract. A Western blot is shown using GATA-1 N6 antibody. (B) Lane 1, 8 μL of anti-GATA-1 N6 and 1 μL of anti-rat IgG2a; lane 2, 8 μL of anti-p53 (PAB 246) and 1 μL of anti-mouse IgG; lane 3, 8 μL of anti-cyclin B1 (GNS1) and 1 μL of anti-mouse IgG for 1.5 hours at 4°C with rotation, and 40 μL of protein G plus agarose was added with 4 more hours of rotation at 4°C; lane 4, MEL nuclear extract. A Western blot with HRP-conjugated anti-p53 is shown.
