Figure 5. Effect of high concentrations of fetuin-A and albumin on the formation of NB-like particles in metastable versus supersaturated medium.

Stock solutions of BSF and/or HSA were diluted into DMEM to the concentrations indicated on the top, so as to mimick the concentration range of these proteins seen in HS. When indicated, the precipitating reagents CaCl₂ and NaH₂PO₄ were added to the concentrations shown at the bottom (labeled as “Calcium+Phosphate Added”). A₆₅₀ turbidity readings were performed following inoculation (“Day 1”) and after incubation in cell culture conditions for “1 Month”, as indicated on the right. Seeding of the proteins alone without additional precipitating reagents did not produce any turbidity increase even following incubation for 1 month (“None” column). Notice the bell-shaped turbidity increases seen with 0.3 mM and 0.5 mM calcium and phosphate columns, observed at “1 Month,” but not “Day 1.”