Figure 6

(A) Percentage of cells with fluorescence values included in the yellow area (or in the gate M1) in the representative flow cytometry 2D and 1D plots reported in the Supplementary Information. Significant difference from control value was indicated by *(p < 0.05) (one-way ANOVA with Tukey’s post hoc test) (B) Cu(II) enhanced the accumulation of ubiquitinated proteins in Hela cells. Hela cells were treated with 20 and 80 μM of Cu(II) for 24 hours, and then ubiquitinated proteins were detected by Western blot (C). Proteasome disassembly following HeLa cells incubation in the absence (lane 3) and with 20 (lane 4) and 80 μM Copper(II) (lane 5). Isolated 20S and 26S proteasomes are also analyzed as a control (lanes 1 and 2). At 24 h of incubation equal numbers of cells were harvested, and lysates were separated by nondenaturing PAGE and proteasome activity was evidenced by in-gel peptidase activity. The proteasome species are indicated as CP-RP₂, doubly capped 26S; CP-RP, singly capped 26S; and CP, uncapped 20S.