Figure 7. Phosphorylation of eIF2α during Mitochondrial Stress Requires ROS.
(A) Fluorescent photomicrographs of clk-1(qm30);hsp-60pr::gfp animals synchronized and raised on control or plates containing 8 mM ascorbate. Images were obtained on day 5. (B) Immunoblot of phosphorylated eIF2α from clk-1(qm30) and isp-1(qm150) mutant worms untreated or treated with 25 mM ascorbate. The anti-HDEL immunoblot serves as a loading control. Worms were synchronized and allowed to develop to adulthood, at which time they were treated with 25 mM ascorbate for 16 hours prior to harvest. (C) Immunoblot of phosphorylated eIF2α from wild-type and gcn-2(ok871) animals treated with 1 mM paraquat (PQ). The anti-HDEL immunoblot serves as a loading control. Worms were synchronized and raised in liquid culture to the young adult stage when 1 mM PQ was added for 16 hours prior to harvest.