Figure 4.

Bax protein levels in parental and LH-RH-Rc-expressing cells. Stable LH-RH-Rc COS cells and parental COS cells were incubated for 36 h in the presence of 10 μM ecdysone analog, followed by administration of AN-201, AN-207, and/or Cetrorelix at doses indicated for 1 h. Equal amounts of protein (70 μg/lane) were separated by 10% SDS/PAGE and transferred onto poly(vinylidene difluoride) membranes. Membranes were incubated with anti-Bax and antiactin antibodies for 1 h at room temperature. Secondary antibodies used were anti-rabbit or anti-mouse IgG conjugated with horseradish peroxidase. Western blots were visualized by chemiluminescence.