The HEK293T cells (CRL-11268, ATCC) were grown at 37 °C and 5% CO2 in a humidified atmosphere incubator (Thermo). The culture medium contained Dulbecco’s modified Eagle’s medium (Gibco), 10% fetal bovine serum, and penicillin-streptomycin (50 μ/ml and 50 μg/ml). The HEK293T cells were seeded at the density of 100,000 per 8-mm × 8-mm, and the cover slip was coated with poly-lysine (Sigma) in a 24-well plate before transduction.
The dissociated hippocampal neurons were prepared from P0 pups, as described previously14 (link). The neurons were plated at the density of 80,000 per 8-mm × 8-mm cover slip coated with poly-lysine (Sigma). The infected neuronal cultures were fixed with 4% paraformaldehyde/4% sucrose in phosphate buffered saline, and fluorescent images were collected with a laser confocal microscope (Olympus) using a 20× objective.
The dissociated hippocampal neurons were prepared from P0 pups, as described previously14 (link). The neurons were plated at the density of 80,000 per 8-mm × 8-mm cover slip coated with poly-lysine (Sigma). The infected neuronal cultures were fixed with 4% paraformaldehyde/4% sucrose in phosphate buffered saline, and fluorescent images were collected with a laser confocal microscope (Olympus) using a 20× objective.
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