Mouse primary hippocampal neurons (mPHN) were isolated from E17 to E18 C57Bl/6J (Janvier) (IMSR Cat# JAX_000664, RRID:IMSR_JAX:000664) embryos, dissociated, and plated in 96-well plates (Greiner, μClear, 655946) previously coated with poly-L-lysine (Sigma Aldrich, P1274) at 20,000 cells per well. Neurons were kept at 37 °C and 5% CO2 in B-27 and GlutaMax supplemented Neurobasal medium (Gibco, 10888022) (76 (link), 77 (link)).
QBI-HEK 293A (RRID:CVCL_6910) cells were cultured in DMEM (Gibco, 11960-044) supplemented with 10% FBS and 1% PenStrep (Gibco, 15140163). In total, 5000 cells were plated per well in 96-well plates and kept at 37 °C and 5% CO2. Forty-eight hours after plating, cells were treated with 1 μM YM-201636 and incubated for 30 min before being fixed with 4% paraformaldehyde and stained for LAMP1 (Abcam, Cat# ab25245, RRID:AB_449893 ) and F-actin (Thermo Fisher, Cat# A22287, RRID:AB_2620155 )
QBI-HEK 293A (RRID:CVCL_6910) cells were cultured in DMEM (Gibco, 11960-044) supplemented with 10% FBS and 1% PenStrep (Gibco, 15140163). In total, 5000 cells were plated per well in 96-well plates and kept at 37 °C and 5% CO2. Forty-eight hours after plating, cells were treated with 1 μM YM-201636 and incubated for 30 min before being fixed with 4% paraformaldehyde and stained for LAMP1 (Abcam, Cat# ab25245, RRID:
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