Igg conjugated alexa fluor 488

Flow cytometry analysis was performed at Day 35 of GABAergic interneuron differentiation in order to assess the purity of the cell population. Cells were dissociated and fixed with 4% paraformaldehyde and stained with the following antibodies: NKX2.1 (sc-13040, Santa Cruz Biotechnology), and GABA (A2052, Sigma). The secondary antibody was IgG-conjugated Alexa Fluor 488 (Life Technologies). Live and dead cells were identified using the Zombie UV Fixable Viability Kit (Biolegend). Unstained as well as IgG-conjugated Alexa Fluor 488 (Life Technologies) secondary antibody alone was used as a control and to gate positive populations. Cells were analyzed on a BD LSR II Flow Cytometer (BD Biosciences), and data were interpreted with FlowJo Version 10.

All neural progenitors, including MGE cells, and further differentiated cells, including MGE cell-derived GABAergic neurons, were fixed in 4% paraformaldehyde and stained with the following primary antibodies: nestin (MAB5326, EMD Millipore), Pax6 (Developmental Studies Hybridoma Bank), FoxG1 (sc-48788, Santa Cruz Biotechnology), MAP2 (MAB3418, AB5622, EMD Millipore), Tuj1 (MAB1637, EMD Millipore; MRB-435P, Covance), apoE (178479, Calbiochem), PHF1 (gift from Peter Davies), AT8 (MN1020, Thermo Fisher Scientific), AT180 (MN1040, Thermo Fisher Scientific), Tau5 (577801, EMD Millipore), total-tau (T6402, Sigma), NKX2.1 (sc-13040, Santa Cruz Biotechnology), GABA (A2052, Sigma), and cleaved Caspase-3 (D3E9, Cell Signaling Technology). The secondary antibodies were IgG-conjugated Alexa Fluor 488 or Alexa Fluor 594 (Life Technologies). Nuclei were stained with DAPI. Images were taken with a Leica epifluorescence microscope, a Keyence BZ-9000E fluorescence microscope, or a Leica confocal imaging system.