(A–C) Total cellular levels (TotCell), survival rate (as in 3B, D), and EGSH.ER values in HGT1 and HGT1-Δsec61-sec61Δ7 cells incubated with 30 μM GSH for the indicated time. (D–G)
EGSH.ER values in HGT1 cells (D–G), HGT1-Δsec61sec61N302L, or HGT1-Δsec61Δssh1sec61N302L as indicated (D), HGT1-Δsec61sec61N302D (E), HGT1-Δsec61Δplug (F, G), incubated during the indicated time with the indicated amount of GSH. (H) Value of EGSH.ER in HGT1-Δerg6Δpdr1Δpdr3 cells after incubation with 50 μM GSH during the indicated time. Puromycin (2 mM) was added 20 min prior to GSH addition. (I) Value of EGSH.ER in HGT1 cells after incubation with 50 μM GSH during the indicated time. Cycloheximide (CHX) (200 μg/mL) or vehicle (DMSO) was added 15 min prior to adding GSH. (J). Value of EGSH.ER in HGT1 cells expressing SRP54 or SRP54dn from a GAL1 promoter incubated 60 min with 30 μM GSH, 3 h after switching the medium from raffinose to galactose. All values are the mean of three independent biological replicas ± standard deviation (s.d.)