Figure 3.

NM-GFP Aggregates Induced by Fibrillized Peptide p103–113 Exhibit Prion Properties

(A) Confocal microscopy analysis of N2a NM-GFP^sol cells 2 days after exposure to fibrillized peptide p103–113. Scale bar, 5 μm.

(B) Schematic diagram of the workflow for isolating N2a NM-GFP cell clones harboring NM-GFP aggregates induced by p103–113 and subsequent coculture. Isolated clones were cocultured with recipient N2a NM-HA^sol cells for 24 h to study the intercellular aggregate transmission efficiency.

(C) Presence of NM-GFP aggregates in cell clones. The N2a NM-GFP^agg cell clones 1 and 2 contain several small NM-GFP aggregates. Nuclei were stained with Hoechst (blue). Scale bar, 5 μm.

(D) N2a NM-GFP^agg donor clones and recipient N2a NM-HA^sol cells were cocultured for 24 h and subsequently assessed for the percentage of N2a NM-HA-expressing cells with induced NM-HA aggregates. Shown is the mean ± SD (n = 3). No aggregates were observed in NM-HA^sol cells that were cultured alone.

(E) Donor N2a NM-GFP^agg clone 2 was cocultured with recipient N2a NM-mCherry^sol cells. Aggregate induction was monitored by time-lapse microscopy for 4 h after co-plating. Images from time points 20–160 min are shown. Arrows indicate aggregates. Scale bar, 5 μm.

See also Video S1.