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. 2009 Mar 13;284(11):6972–6981. doi: 10.1074/jbc.M805956200

FIGURE 5.

FIGURE 5.

Delivery of anti-synuclein antibody H3C into primary neurons. A, fluorescence microscopy images of primary neurons treated with nanoparticles loaded with a monoclonal antibody H3C against α-synuclein or (B) with H3C and Polysorbate 80 alone and stained with a Alexa Fluor 594-conjugated secondary and co-stained with DAPI. C, Western blot analysis of whole-cell lysates (W) and protein-G pulldown assays (G) from neurons treated with H3C-NPs (lanes 3 and 4), empty nanoparticles (lane 5 and 6), or H3C-NPs after preincubation with anti-LDL receptor antibody (lanes 7 and 8). Lane 1 = H3C antibody (50 ng), lane 2 = recombinant human α-synuclein (50 ng). Blot was probed with a goat antibody (SC7012) against N-terminal α-synuclein. D, immunocytochemistry of neurons treated with H3C-NPs then fixed and stained with SC7012, a goat antibody against the N terminus of α-synuclein. The primary antibodies were detected using Alexa Fluor 488-conjugated anti-mouse and Cy3-conjugated anti-goat, respectively. NP-delivered H3C colocalizes with SC7012. Scale bar = 50 μm.