Poly d lysine coated glass coverslips

C57BL/6 transgenic mice expressing human α-Syn (Jackson Laboratory, 017,682) were used for primary cortical neuron isolation. Primary cortical neuronal cells from mice during embryonic days 16–18 were isolated using Accutase solution (Sigma, A6964) and maintained as previously published [21 (link)]. Briefly, neuronal cells were plated on poly-D-lysine-coated glass coverslips (Corning, Inc.) at a density of 2 × 105 cells/mL in a 24-well plate containing neurobasal medium (Gibco, 12,348,017) supplemented with 2% B-27, 0.5 mM GlutaMax (Gibco, 35,050–061), 10,000 units/mL penicillin, 10,000 μg/mL streptomycin, and 25 μg/mL amphotericin B supplement. Media changes were performed every 3–5 days by replacing 50% culture media with fresh media. Cells were grown for 10–13 days in vitro (DIV) before experiments. Primary cortical neurons were grown on coverslips in 24-well plates. α-Syn oligomeric polymorphs (0.5 μM), generated as previously published [21 (link)], were incubated with each SynTC (2 μM) for 30 min at RT. Primary cortical neurons were then exposed for 24 h [44 (link), 45 (link)]. The procedures involving experimentation on animal subjects are done in accordance with UTMB’s guidelines.

The C57BL/6 animals (Jackson Laboratory, 000664) were used for primary cortical neuron isolation. Primary cortical neuronal cells from C57BL/6 mice during embryonic days 16–18 were isolated using Accutase solution (Sigma, A6964) and maintained as described elsewhere [65 (link)]. Neuronal cells were plated on poly-D-lysine-coated glass coverslips (Corning, Inc.) at a density of 2 × 10⁵ cells/mL in a 24-well plate containing neurobasal medium (Gibco, 12348017) supplemented with 2% B-27, 0.5 mM GlutaMax (Gibco, 35050-061), 10,000 units/mL penicillin, 10,000 μg/mL streptomycin, and 25 μg/mL amphotericin B supplement. Half of the Media changes were performed every 3–5 days by replacing 50% culture media with fresh media. Cells were grown for 10–13 days in vitro (DIV) before experiments.