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Fig. 7 | Molecular Brain

Fig. 7

From: Postsynaptic density protein 95 (PSD-95) is transported by KIF5 to dendritic regions

Fig. 7

Complexes of PSD-95-KIF5 colocalized with GluA1 particles in dendrites. Cultured hippocampal neurons were transfected with PSD-95-GFP constructs and incubated for days. The cultures were immunostained with monoclonal anti-PSD-95 antibody and polyclonal GluA1 antibody; they were subsequently stained with C3-conjugated anti-mouse IgG and Alexa−Fluor® 647 anti-rabbit IgG antibody. a Representative images of immunostaining in the first row. Each image was merged to show colocalization in the second row. A colocalized image of PSD-95 and KIF5 was collated with the image of GluA1. The colocalized points appeared white. Scale bar: 20 μm. b Boxed dendrites were enlarged to see the colocalization of GluA1 with the complex of PSD-95-KIF5A. c Staufen expression increased the association of PSD-95 and KIF5A. HA-PSD-95 and FLAG- KIF5A were cotransfected with 1, 2, or 3 μg of Myc-Staufen, or without Myc-Staufen as a control. After immunoprecipitation using anti-FLAG antibody or mouse IgG, immunoprecipitates were analyzed by Western blotting using anti-HA antibody. The lower panel shows expression of each group. d Quantified data of Western blot analyses (0 μg of Staufen: 100.0% ± 0.00%, n = 3; 1 μg of Staufen: 141.6% ± 16.96%, n = 3; 2 μg of Staufen: 192.3% ± 6.59%; 3 μg of Staufen; 274.4% ± 42.30%, n = 3). N values indicate the number of independent experiments. e Schematic diagram showing GluA1-containing vesicle transport mediated by PSD-95-KIF5A complex in dendrites. TARP: transmembrane AMPA receptor regulatory protein

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