FIGURE 3.

BAX and IGFBP-3 interaction in dot blot experiments. A, the peptides used for dot blots against anti-IGFBP-3 included HN (a known binding partner of IGFBP-3 used as positive control, 2, 5, and 10 nmol), BCL-2 (fragment, 1, 2, and 5 μg), BAX (mouse full long sequence, 1, 2, and 5 μg), and BSA (used as negative control, 1, 2, and 5 μg), IGFBP-3 (used here as positive control for anti-IGFBP-3 antibody, 1, 2, and 5 μg). B, the reverse experiment protein dots versus with anti-BAX were: HN (a known binding partner of BAX used here as positive control, 2, 5, and 10 nmol), BSA (used here as negative control, 1, 2, and 5 μg); IGFBP-3 (1, 2, and 5 μg); HN mutant (HN-C8P, a non-BAX-binding mutant; 2, 5, and 10 nmol), and BAX (used here as positive control for anti-BAX antibody, 1, 2, and 5 μg). Both dot blots confirmed the interaction between BAX and IGFBP-3.