Fig 5.

Ino80, SRCAP, and URI are not essential for type I IFN-stimulated transcription. (A and B) Nuclear proteins prepared from HEK293 cells transfected with the indicated siRNAs were immunoblotted for Ino80 (A) or SRCAP (B) and HDAC2 (loading control). (C) HEK293 cells carrying two independent inducible hairpins against URI were treated with doxycycline for 72 h or left untreated. The lysates were blotted for URI and HDAC2. (D) Cells were transfected with the indicated siRNA duplexes and treated with IFN-α2a for 8 h before isolation of RNA. Expression of ISG56, IFI6, 9-27, and actin mRNAs was quantified by real-time PCR and is presented as the fold induction over uninduced levels. The averages for three experiments are shown. (E) The HEK293 cells described for panel C were incubated with IFN-α2a for 8 h before RNA isolation. Expression of ISG56, IFI6, and actin was quantified by real-time PCR and is presented as the fold induction over uninduced levels before and after URI depletion. The averages for three experiments are shown. (F) HEK293 cells were infected with the indicated hairpin, transfected with the indicated siRNA duplexes, and treated with IFN-α2a for 8 h before isolation of RNA. Expression of ISG56, IFI6, and actin mRNAs was quantified by real-time PCR and is presented as the fold induction over uninduced levels. The averages for three experiments are shown.