Expression Nsp6 orthologs does not inhibit mTOR or induce CHOP. HEK cells expressing the nsp6 orthologs were lysed and analyzed by SDS PAGE followed by protein gel blot. (A) In parallel experiments HEK cells were transfected with (Lane 1) IBVnsp4mCherry, (Lane 2) IBVnsp6mCherry, (Lane 3) SARSnsp6mCherry, (Lane 4) MHVnsp6mCherry, (Lane 5) PRRSVnsp5–7mCherry, (Lane 6) left untreated, (Lane 7) incubated with thapsigargin to induce ER stress, (Lane 8) Torin-1 to inhibit mTOR, or (Lane 9) starved in HBSS to induce autophagy, as indicated. Protein gel blots were probed with antibodies to actin as a loading control, anti phospho4E-BP1 and CHOP. (B) Shows expression of transfected proteins at x40 magnification; (1) IBVnsp4mCherry, (2) IBVnsp6mCherry, (3) SARSnsp6mCherry, (4) MHVnsp6mCherry, (5) PRRSVnsp5–7mCherry. (C) Transfected cells, cells incubated with thapsigargin and control HEK293 cells were subjected to PCR analysis using primers specific for XBP1 mRNA.