FIG 3.

Insensitivity of mTORC1 to AA withdrawal in HSV-1-infected cells is controlled in part by TSC. Growth-arrested NHDFs treated with TSC2 siRNA (siTSC2) or control, nonsilencing siRNA (NS) were infected with the F strain ΔUs3 mutant (ΔUs3-F) (A) or mock infected (B). At 12 hpi, cultures were treated with Akt VIII, and at 15 hpi, they were either incubated in AA-free RPMI 1640 (−) for 50 min, incubated in AA-free RPMI 1640 for 50 min followed by a 30-min stimulation in AA-replete RPMI 1640 (±), or left in AA-replete DMEM for 50 min (+). Total protein was isolated, fractionated by SDS-PAGE, and analyzed by immunoblotting using the indicated antibodies. The α subunit of eukaryotic initiation factor 2 (eIF2α) and HSC70 served as loading controls for panels A and B, respectively.