Fig 2. CHIP-T246M has chaperone activity in cell-free assays.

(A) Relative luciferase activity on heat-denatured luciferase after one hour incubation with the indicated CHIP proteins (CTRL = no protein, IgG = immunoglobulin G), either in the absence (left) or presence of HSP40 and HSP70, represented by dot plot and summarized by the mean ± 95% CI, N = 4 experimental replicates: *, **** correspond to p < 0.05, and 0.0001 via Tukey’s post hoc test compared to CHIP-WT, †, †††† correspond to p < 0.05, and 0.0001 via Tukey’s post hoc test compared to IgG; ns indicates p > 0.05 compared to IgG. (B) AMPK kinase activity towards the Z’LYTE peptide substrate in the presence of increasing amounts of the indicated recombinant CHIP proteins or IgG protein control represented by scatter plot and summarized by the mean ± 95% CI, N = 3 experimental replicates. The half maximal effective concentration (EC₅₀) for each protein is included.