Fig. 2.

One to two days of incubation with AβOs impaired LTP-induced increase in the amount of GluA1-SEP in either PSLM or non-PSLM. (A) Schemes of PSLM and live-cell imaging of GluA1 or GluA2 labeled with SEP (GluA-SEP). While GluA-SEP is fluorescent on the cell surface, the fluorescence is quenched in cytoplasmic vesicles at a low pH. TIRF illumination activates fluorescent molecules within the limited Z-axis depth (about 100 nm), enabling high signal-to-noise detection of signals. (B) Representative images (left) of the GluA1-SEP signal (green) and PSD95-RFPt signal (magenta). PSD95-RFPt was recorded before the stimulation and merged with the time-lapse images of GluA1-SEP. In neurons treated with revAβ, the GluA1 signal increased in PSLM (white arrows) and non-PSLM (white arrowheads). Averaged time courses (right) of GluA1-SEP fluorescence intensity in PSLM (red) and non-PSLM (black) measured every 20 sec before and after the LTP stimulation (black arrows). (C) Statistical analyses of data shown in (B). Averaged fluorescence intensity at each time point in 3 min bin. Significant differences between revAβ and AβOs (Benjamin-Hochberg test), or before and after the stimulation (Dunnett's test) are indicated by asterisks (N = 18, 19 cells, *P < .05, **P < .01).