Extended Data Fig. 2: Intraneuronal APP-βCTF/Aβ accumulates selectively within pa-AL in AD mice.
a. AV fractionation from 10-month-old Tg2576 mice. Fractions were validated by organelle markers (lysosome: CTSD, mitochondria: Tom20, ER: SEC61B, AV: p62) and anti-Aβ antibody 4G8. Experiment was repeated 2 times independently with similar results. b. Immunofluorescence co-labeling of 5-month-old Tg2576/TRGL mouse brain neurons with an antibody against Aβ1−42 (JRF/cAβ42/26). Aβ accumulates in enlarged pa-AL producing a white signal (arrowhead). Experiment was repeated 3 times independently with similar results. Scale bar 20 μm. (c) Quantitation graph of the PLA fluorescence per neuron from N2A-APPswe cell (N2a (0.9±0.2), N2a APPswe (19.6±1.1)) and (d) 10-month-old Tg2576 mouse brain compared with WT controls. WT (1.4±0.1), Tg2576 (6.9±0.5). n=50 cells per each. e-g, Quantitation graph of the PLA fluorescence per neuron from 10-month-old Tg2576/TRGL. e. Total number of PLA signal per neuron. TRGL (1.2±0.1), Tg2576/TRGL (6.2±0.3). (f) Number of the PLA signal in pa-AL per neuron. TRGL-pa-AL (0.1±0.0), Tg2576/TRGL-pa-AL (5.8±0.2). (g) Percentage of PLA signal in pa-AL in neuron compared with WT controls. TRGL-pa-AL (6.7±3.4 %), Tg2576/TRGL-pa-AL (92.9±1.3 %). n=50 cells. Quantitative data are presented as means ±S.E.M. unpaired t-test, two-tailed P value as indicated. a-d: Experiment was repeated 3 times independently with similar results.
