A novel fission yeast gene, tht1+, is required for the fusion of nuclear envelopes during karyogamy - PubMed (original) (raw)

Figure 11

Protease protection assay. Microsome fractions prepared from conjugating cells carrying the plasmids pNT75 (Tht1-NcoI-HA), pNT74 (Tht1-StuI-HA), pNT28 (Tht1p), or pKD10 (vector) were treated with trypsin either in the absence or presence of Triton X-100 and subjected to Western immunoblot analysis. (A) Western blots probed with anti–HA antibodies. The amount of trypsin added was 0 μg/ml (lanes 1, 6, 11, and 16), 0.5 μg/ml (lanes 2, 7, 12, and 17), 1.5 μg/ ml (lanes 3, 8, 13, and 18), 3 μg/ml (lanes 4, 9, 14, and 19), and 5 μg/ml (lanes 5, 10, 15, and 20). (B) Western blots probed with anti–Tht1 antibody, 1-3-2. The amount of trypsin added was 0 μg/ml (lanes 1, 4, 7, 10, 13, 16, and 19), 0.5 μg/ml (lanes 2, 5, 8, 11, 14, and 17), and 5 μg/ml (lanes 3, 6, 9, 12, 15, and 18). The open arrow indicates a nonspecific band.