Fig. 3.

Inhibition of nitrite reduction abolishes coclustering of HS-anMan and Aβ and increases SYNfil formation during growth in the presence of the β-secretase inhibitor. Representative immunofluorescence images of cells that were grown to confluence in A, D) regular medium (UT, untreated), or in B, E) medium containing 100 nM of β-inhibitor (LY2811376), or in C, F) medium containing 100 nM β-inhibitor (LY2811376), 150 μM allopurinol, and 1 μM SB203580. Staining was performed with mAb AM (for HS-anMan, green), mAb SYNfil (green), pAb Aβ40 (for the Aβ region, red), and DAPI (for nuclei, blue). Exposure time was the same in all cases. Bar, 20 μm. Inserts, immunoblots of RIPA extracts of cells that were grown to confluence in regular medium (UT) or in medium containing the β-secretase inhibitor or in medium containing both the β-secretase inhibitor and the nitrite reductase inhibitors. The extracts were slot blotted to PVDF membranes which were probed with A–C) mAb AM, or D–F) mAb SYNfil. The error bars indicate SE.