Figure 3.

Knock down of Atf6 inhibits TOR signaling. S2R+ cells were transfected with luciferase (Luc), TSC1, eIF2a, IRE1, PERK, or Atf6 double-stranded (ds) RNA. The transfected cells were cultured for 2 days, and incubated with serum-free media for 1 hour before cells were collected. Then, Western blot analysis was performed with phospho(Thr398)-specific S6K antibody or tubulin antibody. Firefly luciferase (Luc), Tuberous sclerosis complex 1 (TSC1), Eukaryotic initiation factor 2a (eIF2a), Inositol requiring enzyme 1 (IRE1), Pancreatic ER kinase (PERK), and Activating transcription factor 6 (Atf6).