FIGURE 1.

Shotgun analysis of the secretome of primary neurons after BACE1 inhibitor treatment and sample quality control. A, scheme summarizing preparation of neuronal culture samples and shotgun analysis. B and C, quality control Western blots indicating specific inhibition of cleavage of APP by BACE1 in neuronal cultures after β-secretase inhibitor IV (BI-IV)(1 μm) treatment. The cell lysates (B) were separated on 16% Tricine gel and blotted with antibodies B63 (against APP C terminus), D10E5 (BACE1), and anti-β-actin. The conditioned media (C) were concentrated and separated on 4–12% Tris-SDS-PAGE and blotted with antibodies anti-sAPPβ and 22C11 (against the N terminus of APP).