Fig. 2

Aβ treatment increases the interaction between PRAK and RAGE. a Aβ treatment increases RAGE-PRAK binding. Overexpression of RAGE and PRAK in SH-SY5Y cells were treated with either DMSO or monomeric Aβ 2 μM for 6 h and co-localization was detected using in situ proximity ligation assay kit. Probes in close proximity (<40 nm) are indicated by fluorescent dot signals. Scale bars represent 20 μm. b Aβ enhances the co-localization between RAGE and PRAK. SH-SY5Y cells was induced by a 6 h treatment of DMSO or monomeric 2 μM Aβ. Immunofluorescence analysis using anti-PRAK (green) and anti-RAGE (red) antibodies. SIM was used for image analysis. Scale bars represent 5 μm. c PRAK and RAGE binding in vivo. Immunoprecipitation (IP) with anti-RAGE antibody was accomplished using total brain lysates from 3 months wild-type litter mates and Tg6799, followed by western blotting with anti-PRAK antibody. Quantification of protein level was performed by densitometric analysis. Data are mean ± SEM. n = 4. *P < 0.05