Fig. 1: RSL3-induced calibration of the UPS is distinct from proteasome inhibition.
From: Activating the NFE2L1-ubiquitin-proteasome system by DDI2 protects from ferroptosis
a Immunoblot of ubiquitin from EA.hy926 cells treated with 5 µM RSL3 and 100 nM bortezomib (BTZ) for 9 h and 3 h, respectively. b Immunoblot of NFE2L1 in EA.hy926 cells treated with indicated time points and concentrations of RSL3 (FL: full-length form ca. 120 kD; CL: cleaved ca. 95 kD). c Immunoblot of NFE2L1 in nuclear and cytoplasmic fractions isolated from EA.hy926 cells treated as indicated for 3 h. d Native page of EA.hy926 cells treated with 5 µM RSL3 and 100 nM BTZ for 6 h with in-gel activity and immunoblot of the ɑ1-7 (20S) subunits. e Proteasomal activity in EA.hy926 cells treated with 5 µM RSL3 for 3 h. f Proteasomal activity assay in EA.hy926 cells with the extracts being incubated with indicated concentrations and time points of RSL3 and proteasome inhibitor MG132. g Volcano Plot of the ubiquitome of EA.hy926 cells treated for 9 h with 5 µM RSL3 Padj < 0.05 indicated in red. (n = 4 replicates). h Principal component analysis (PCA) of the ubiquitome of EA.hy926 cells treated with 5 µM RSL3 and 100 nM BTZ (n = 4 replicates). i Top 10 enriched gene ontology (GO) terms in the ubiquitome of EA.hy926 cells treated for 9 h with 5 µM RSL3.
