Extended Data Fig. 9: Glucose starvation does not cause ribosome collisions.
From: The atypical E3 ligase HOIL-1 safeguards the ribosome during cellular stress
a, Polysome profiles from lysates of untreated AC16 cells. b, Polysome profiles from lysates of AC16 cells treated with anisomycin (5 µM) for 1 hour. c, Polysome profiles from lysates of AC16 cells starved of glucose for 24 h. d, Immunoblot analysis of sucrose fractions from a-c. e, Schematic of ribosome stalling reporter constructs. Ribosome stalling on (AAA)20 reduces RFP translation. f, Flow cytometric analysis of GFP and RFP in AC16 cells stably expressing the ribosome stalling reporter transfected with ZNF598 or NTC siRNA 48 h prior (n = 4 biological replicates). Immunoblot analysis of ZNF598 knockdown shown. g, Flow cytometric analysis of GFP and RFP in AC16 cells stably expressing the ribosome nonstalling reporter (n = 3 biological replicates). h, Flow cytometric analysis of GFP and RFP in AC16 cells stably expressing the ribosome stalling reporter (n = 3 biological replicates). Data are presented as mean ± s.e.m from one representative experiment. P values were determined by unpaired two-tailed Student’s t-tests (f, h). Source numerical data and uncropped blots are available in Source Data.
