Deducing the mode of hV3 aggregation. (A) Schematic of how cooperativity of unfolding and aggregation are measured as ΔH_ME-app (left) and ΔT_m (right), with values normalized between 0 and 1. (B) Summary of how the kinetics of unfolding and aggregation can be inferred from ΔH_ME-app and ΔT_m. Comparison of ΔH_ME-app and ΔT_m is useful to study the aggregation propensity and rate of aggregation of proteins. A high or low value of various parameters indicates unfolding, aggregation, or unfolding coupled with aggregation. (C) Comparison of ΔH_ME-app and ΔT_m calculated from ME₂₁₅. Here, the most stabilizing and destabilizing mutants derived based on the global comparison of ΔH_ME-app and ΔT_m are shown, as described in Fig. S8. Data for WT is shown in gray for comparison only. Error bars represent SD derived from a minimum of three independent experiments. Statistical significance (***, P < 0.001; **, P < 0.01) was derived using t test (unpaired). See Figs. S7 and S8 for the complete data and comparison using the heat map scheme.