Figure 1. BAG3 is induced upon proteasome inhibition and forms cytoplasmic puncta. (A–C) HEK293T cells were either left untreated or treated with 20 μM MG132 for 5 h or 100 nM bortezomib overnight. Total proteins (A and B) or mRNA (C) were extracted and BAG3 or BAG1 protein or BAG3 mRNA levels were measured (*P < 0.05 compared with control; n > 3 independent samples ± sem). (D–H) HeLa cells were treated for 3 h with 20 μM MG132 and cells were fixed with formaldehyde followed by acetone treatment. Subcellular distribution of endogenous BAG3, HSPA1A, SQSTM1, and ubiquitin was investigated by immunofluorescence using specific antibodies.