Figure 3

Immunostaining of acidic vesicles and p62 or LC3 under inflammatory stress in primary tri-cultures. Co-labeling of autophagic receptor (A) p62 (green) or (B) LC3 (green), Lyso-ID for acidic vesicles (red), and DAPI for nuclei (cyan) in tri-cultures seeded on coverslips, exposed to 20 μM Aβ42 or LPS 100 ng/mL in serum-free medium for 48 hours. All images were from a compilation of the entire z-series sections acquired by confocal microscopy (Olympus IX-81). A white square represents a magnified ROI. Two-dimensional fluorograms display the intensity and distribution of different colored pixels within the magnified ROI in the merged image. Scale bars, 42 μm. DAPI, 4′,6-diamidino-2-phenylindole; LPS, lipopolysaccharide; ROI, region of interest.