Effect of Crm1 inhibition/depletion on Nup2 binding patterns and of Nup2 degradation on Crm1 binding patterns in budding yeast

We found that Crm1 and Nup2 bind at similar sites in the genome. To investigate their interdependence and to see if one of the two proteins is functioning upstream, we depleted one protein while monitoring changes in binding pattern of the other. Yeast cells were grown in synthetic complete medium containing glucose (SDC) at 30C. Crm1 was inhibited or depleted by treating yeast cultures with 185 nM leptomycin B (LMB) in ethanol for 30 min or with 1 uM estradiol (E2) in ethanol for 30 min followed by auxin (5-Ph-IAA) for 2 hours at 30C. Nup2 was degraded by treating cell cultures with indole-3-acetic acid (3-IAA) in ethanol for 16 hours at 30C. After treatment, ChEC-seq2 was performed for Nup2 or Crm1. All experiments were performed in biological triplicate.