Figure 7. Seeding of NB-like particles by boiled fetuin-A and albumin in metastable versus supersaturated medium.

(A) Metastable medium: BSF and bovine serum albumin (BSA) were boiled for the time indicated on the left side. The boiled protein solutions were then diluted into DMEM, separately or together, to the final concentrations indicated on the top. A₆₅₀ was then monitored following inoculation (“Day 1”) and after incubation in cell culture conditions for 1 month, as indicated on the right. Inoculation of either untreated BSF or BSF that had been boiled for either 10, 30, or 120 min did not result in any turbidity changes after 1 month of incubation. In contrast, pre-boiled BSA produced either bell-shaped or linear turbidity changes depending on the amount of boiling time. When added together, pre-boiled BSF and BSA produced additive effects. (B) Supersaturated medium: BSF and BSA were added to medium as in (A) that was also inoculated with 1 mM each of CaCl₂ and NaH₂PO₄. Notice the various patterns of turbidity changes associated with the three protein combinations. See the text for further details.