Figure 6.

PIDD requires Hsp90 for its functional activity. (a) HEK293T wild-type cells were pretreated with 1 μM GA for 20 min (etoposide treatment) or for 60 min (TNFα treatment) in presence of 10 μM MG132. Etoposide (40 μM) was added for 1 h and TNFα (10 ng/ml) for 10 min. Lysates were analysed by western blotting. (b) HEK293T cells were pretreated with 1 μM GA for 20 min. Etoposide (40 μM) was then added for the indicated times. An anti-NEMO immunoprecipitation was performed and analysed by western blotting. (c) PIDD expression was induced in HEK293T Flp-In Trex cells with 0.1 μg/ml doxycycline for the indicated times in presence or absence of 1 μM GA and an anti-Flag immunoprecipitation was carried out. Analysis was performed by western blotting. Molecular weight masses are indicated in kDa and non-specific bands are marked by the asterisk