Fig. 8. Nystatin, filipin and amphotericin B inhibit Sec17p release from vacuoles. Samples equivalent to three standard fusion reactions were incubated at 27°C with Sec18p (13.3 µg/ml), apyrase (0.033 U/ml), anti-Sec18 antibody (66.7 µg/ml), nystatin (50 µg/ml), filipin (200 µg/ml) and amphotericin B (25 µg/ml), where indicated. After the incubation times, vacuoles were removed by centrifugation and samples of the supernatant were analyzed by immunoblotting with anti-Sec17p antibody.