Figure 3.

Hsp90 regulates the constitutive processing of PIDD. (a) Expression of PIDD in HEK293T Flp-In T-Rex cells was induced by 0.1 μg/ml doxycycline in presence or absence of 1 μM GA for 8 h and analysed by western blotting. (b) HEK293T cells stably expressing PIDD were treated with CHX (10 μg/ml), GA (1 μM) or MG132 (10 μM) for the indicated times and the presence of PIDD fragments was monitored by western blotting. (c) Expression of PIDD F445H in HEK293T Flp-In T-Rex cells was induced by 0.1 μg/ml doxycycline for 5 h in the presence or absence of GA (1 μM). PIDD was immunoprecipitated, eluted from the beads and treated with hydroxylamine to induce PIDD processing, which was analysed by western blotting. Blots were exposed simultaneously for the same amount of time. Molecular weight masses are indicated in kDa