Figure 4.

UBR7 could be a proteotoxic stress-specific N-recognin E3 ligase

(A) 200 nM bafilomycin A1, which blocks autophagic flux, is applied for 6 h, and 1 uM MG132 and 200 nM thapsigargin co-treatment, which induces misfolded protein frequency, is applied for 20 h.

(B) UBR7-V5 constructs are transiently expressed and, following stressors, are applied for 24 h except 10 uM of MG132; 25 nM hydroxy chloroquine (HCQ) or 200 nM Bafilomycin A1 for autophagic flux inhibition; 200 μM CoCl₂ for oxidative stress induction; 1 μM MG132 and 200 nM thapsigargin co-treatment; and 1.5 μM geldanamycin treatment for misfolded protein inducing stress. Arrowhead indicates X-nsp4^FLAG. Experiments were performed in duplicates with similar results, and representative blots are shown.