Fig. 2.

Cap-independent RAN translation is upregulated upon stress. a Schematic timeline of stress stimuli and induction of luciferase reporters in HeLa Flp-In cells. b Expression of bicistronic luciferase reporters was induced by doxycycline for 6 h in HeLa Flip-In cells. At the same time, cells were treated with arsenite or MG132 stimuli. Relative expression of AUG-FLuc and C9R-NLuc reporters was compared with no stress control. The luciferase signals were normalized to the total protein amount. Data are mean ± SEM from three biological replicates. ***P < 0.0005, two-tailed t-test. c Stress granules were detected with G3BP immunofluorescence in reporter cells treated with arsenite and MG132. d Immunoblotting using antibody recognizing phospho-eIF2α at the Ser51 site showed the upregulation under stress. GAPDH and total eIF2α were blotted as internal control