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. 2015 Nov 12;291(1):89–102. doi: 10.1074/jbc.M115.680918

FIGURE 2.

FIGURE 2.

Synthetic Fabs inhibit BAX activation induced by pro-apoptotic tBID in liposomal assays. A, synthetic BAX-binding Fabs and a VEGF-specific Fab (YADS1) at 2 μm have no capacity to induce BAX-mediated liposomal ANT/DPX release, whereas tBID induced potent BAX-mediated liposomal release. B, BAX-binding Fabs at 2 μm, except 2A5 and YADS1, inhibit tBID-induced BAX-mediated liposomal ANTS/DPX release. Error bars represent mean ± S.D. at 90 min normalized to tBID-induced maximum ANTS/DPX release at 90 min. C–F, representative liposomal ANTS/DPX release experiments in kinetic representation showing the inhibitory activity of Fabs 3G11, 2B1, and 3E8 at 0.5, 1, and 2 μm Fab. 2A5 had no inhibitory effect. A–F, the experiments were performed with 400 nm BAX, 30 nm tBID, and up to 2 μm Fabs. Data represent mean ± S.D. from triplicates normalized using either tBid-induced BAX mediated ANTS/DPX release as 100% (A and B) or 1% Triton release as 100% (C–F). Data shown are representative of at least three independent experiments.