Chaperone-like activity during heat-induced aggregation of substrates, CS and MDH. (A) Heat-induced CS (2.5 μM) aggregation in the presence of 14.9, 22.5, 29.9 nM B1NTR-NP or PEG-NP (7.48 nM) at 45°C. (B) Heat-induced MDH aggregation in the presence of 7.5, 14.9, 22.5, 29.9 nM B1NTR-NP or PEG-NP (7.48 nM) at 45°C. Each curve is an average of ≥3 replicates. (C) Mixed samples of CS (2.5 μM) and B1NTR-NP were separated into soluble (s) and insoluble (p) fractions and analyzed by SDS-PAGE after shaking for 1 h at 45°C. Samples are: (L) Molecular mass marker, (1) CS (only), (2) CS: B1NTR-NP (14.9 nM), (3) CS: B1NTR-NP (22.5 nM), (4) CS:B1NTR-NP (29.9 nM). (D) Mixed samples of MDH(2.5 μM) and B1NTR-NP were separated into soluble (s) and insoluble (p) fractions and analyzed by SDS-PAGE after shaking for 1 h at 45°C. All SDS-PAGE gels were developed using silver staining. Gel samples are: (L) Molecular mass marker, (1) MDH (only), (2) MDH: B1NTR-NP (7.5 nM),(3) MDH: B1NTR-NP (14.8), (X) spillover lane (4) MDH: B1NTR-NR (29.9 nM)