Journal: Scientific Reports

Article Title: Live Neuron High-Content Screening Reveals Synaptotoxic Activity in Alzheimer Mouse Model Homogenates

doi: 10.1038/s41598-020-60118-y

Figure Lengend Snippet: Validation of PSD95-mVenus postsynaptic, VAMP2-mRFP presynaptic, and colocalized puncta. The majority of anti-VAMP2 and anti-Synapsin1 stained puncta were colocalized with endogenous VAMP2-mRFP puncta. Similarly, the majority of the antibody stained anti-PSD95 and anti-Homer2 puncta were colocalized with endogenous PSD95-mVenus puncta. ( a–c ) Presynaptic VAMP2-mRPF in singly transgenic mice was validated with immunocytochemistry stained ( a ) anti-VAMP2 and ( b ) anti-Syn1, ( c ) 92.1 ± 4.8% of stained VAMP2 puncta and 80.8 ± 3.9% of stained Syn1 puncta was colocalized with endogenous VAMP2-mRFP; ( d–f ) Postsynaptic PSD95-mVenus expression in singly transgenic mice was validated with immunocytochemistry stained ( d ) anti-PSD95 and ( e ) anti-Homer2, ( f ) 107.5 ± 4.5% of stained PSD95 puncta and 113.7 ± 6.2% of stained Homer2 puncta was colocalized with endogenous PSD95-mVenus (mean ± SD%, n = 3).

Article Snippet: All cells were blocked with 5% bovine serum albumin and 0.2% Triton X-100 (Sigma-Aldrich) in PBS for 1 hour at room temperature, and then incubated with anti-Synaptobrevin 2, anti-Synapsin1, anti-PSD95, or anti-Homer2 (Synaptic Systems) antibodies at 1:500 in blocking solution overnight at 4 °C, subsequently washed in 0.1% Triton X-100 in PBS three times.

Techniques: Staining, Transgenic Assay, Immunocytochemistry, Expressing