Extended Data Fig. 5: ATF4 post-transcriptionally regulates NRF2.
From: The atypical E3 ligase HOIL-1 safeguards the ribosome during cellular stress
a, Cell death in 786-O cells transfected with xCT or NTC siRNA prior to glucose starvation for 48 h (n = 4 biological replicates). b, Immunoblot analysis of 786-O cells transfected with xCT or NTC siRNA prior to glucose starvation. c, NRF2 (NFEL2) mRNA levels (normalized to TBP) in AC16 cells after 24 h glucose starvation. Data represented as the fold change over full media (n = 3 biological replicates). d, Immunoblot analysis of AC16 cells transfected with ATF4, NRF2, or non-targeting control (NTC) siRNA 48 h prior to treatment with KI696 (NRF2 activator, 1 µM) for 24 h or sodium arsenite (10 µM) for 8 h. e, Immunoblot analysis of AC16 cells transfected with CHAC1 or NTC siRNA prior to glucose starvation for 24 h. f, Immunoblot analysis of puromycin incorporation in AC16 cells treated with anisomycin (5 µM) or starved of glucose for the indicated times. g, Alexa Fluor 647-o-propargyl-puromycin incorporation in AC16 cells starved of glucose (24 h) or treated with anisomycin (15 min) measured by flow cytometry (n = 3 biological replicates for all except -glucose, n = 4 biological replicates). Data are presented as mean ± s.e.m from one representative experiment (a, c, g). P values were determined by unpaired two-tailed Student’s t-test (a). Source numerical data and uncropped blots are available in Source Data.
