Extended Data Figure 7 : Immobilized NPL4 forms insoluble protein aggregates.

From: Alcohol-abuse drug disulfiram targets cancer via p97 segregase adaptor NPL4

Extended Data Figure 7

a, NPL4–GFP aggregates induced by CuET treatment (1 μM, 3 h) do not co-localize with nuclear speckles (stained by SC-35 antibody) or nucleoli (visible as a DAPI nuclear signal). b, NPL4–GFP nuclear aggregates induced by CuET (1 μM, 3 h) are excluded from chromatin in early prometaphase U2OS cells. c, Co-localization of spontaneous NPL4(MUT)–GFP aggregates with SUMO2/3, poly-UB(K48) and TDP43 in pre-extracted U2OS cells. d, NPL4–GFP aggregates are formed independently of ubiquitylation, as shown in CuET-treated (1 μM, 3 h) cells pre-treated with a chemical UBA1 inhibitor (MLN7243, 10 μM, 1 h). The lack of cellular FK2 staining of ubiquitylated proteins validates the efficacy of the MLN7243 inhibitor. e, Co-localization of FK2 signal with the spontaneous NPL4(MUT)–GFP aggregates in pre-extracted U2OS cells. f, Analysis of p97 in CuET-induced (1 μM, 3 h) NPL4–GFP aggregates in pre-extracted U2OS cells. g, Analysis of p97 in spontaneous NPL4(MUT)–GFP aggregates in pre-extracted U2OS cells. ag, Data are representative of two independent biological experiments.