Extended Data Fig. 8: Effects of perturbing G3P levels on metabolome of senescent cells and characterization of the pEtN synthesis pathway.
A: Heat map showing modules of metabolites in WI38 fibroblast proliferating or infected with a GFP- or GK-overexpressing adenovirus for 7 days. B: Heat map showing modules of metabolites in WI38 fibroblast proliferating or undergoing RAS-OIS and infected with a GFP- or G3PP-overexpressing adenovirus for 7 days. For both panels, a hierarchical clustering approach was used. Data are expressed as row Z scores collected from three biologically independent experiments per condition. C: Measurement of labelled Etn uptake in WI38 fibroblasts proliferating, undergoing DDIS or RAS-OIS for 7 days after a pulse of 1 hour. The reported values are relative to those of proliferating cells. Bars represent the mean of 3 biological replicates +/- s.d. Indicated p values were calculated using an unpaired two-sided Student’s t-test. D: Curves of decay of labelled Etn in WI38 fibroblasts proliferating or undergoing DDIS, after a pulse of 1 hr followed by a chase for the indicated times. For each treatment the values are normalized to those of the 0 hr chase time. Each point represents the mean of 3 biological replicates +/- s.d. Indicated p values were calculated using an unpaired two-sided Student’s t-test. E: Maximal fold change of PCYT2 mRNA in cells induced to senesce under the indicated conditions relative to proliferating cells as measured by Affymetrix microarrays (n = 2 for each sample). F: Representative Western blots showing the indicated protein levels in WI38 fibroblasts proliferating (Prolif.), undergoing RAS-OIS or DDIS for 7 days. The experiment was repeated independently 3 times with similar results.
