Fig 6. SINE RNA promotes IKKβ-mediated phosphorylation of RTA to potentiate viral transcription.

(A) NIH3T3 cells were transfected with FLAG-RTA and either a control construct, B1 SINE, or B2 SINE expression plasmid. 24 h later cells were labeled with [³²P]-orthophosphoric acid for 6 h. Whole cell lysates were precipitated with anti-FLAG antibody and analyzed by autoradiography (top) or western blot using an anti-FLAG antibody (bottom). (B) FLAG-RTA was transfected in to NIH3T3 cells. 24 h later cells were transfected with indicated ASOs and subsequently infected with MHV68 for 6 h in the presence of [³²P]-orthophosphoric acid. Whole cell lysates were precipitated with anti-FLAG antibody and analyzed by autoradiography (top) or western blot using an anti-FLAG antibody (bottom). (C) NIH3T3 cells transfected with the indicated viral promoter luciferase plasmids were cotransfected with wild-type RTA or RTA TTS/A, and control, B1 SINE, or B2 SINE expression constructs. 48 h later luciferase levels were monitored.