Increased eIF2α Phosphorylation and ATF-5 Expression by gfat-1 Overexpression in C. elegans
(A) Representative image of gfat-1 OE strain (gfat-1P::CFP::gfat-1) at late L4 larval stage with cloning scheme of gfat-1 OE construct.
(B) Representative western blot showing phospho-eIF2α levels in gfat-1P::gfat-1 OE strain relative to wild-type controls at day 1 adult stage.
(C) Quantification of western blots. Mean + SD (n = 5), *p < 0.05 (t test).
(D) Western blot showing phospho-eIF2α levels in wild-type animals treated with 10 mM GlcNAc or D-Arg control for 12 h at day 1 adult stage.
(E) Quantification of western blot in (D). Mean + SD (n = 3), *p < 0.05 (t test).
(F) Representative images of atf-5P::gfp reporter strain in gfat-1P::gfat-1 OE background and WT control at day 1 adult stage. Area marked by red dashed line indicates quantified worm area, excluded is the signal from pharyngeal co-injection marker myo-2::mcherry.
(G) Quantification of GFP expression controlled by atf-5 promoter from microscopy pictures at day 1 adult stage (Figure 4F). Mean + SEM (n = 3, 10 animals per condition and experiment), *p < 0.05 (t test).