Chunk #27 — Methods — Immunofluorescence experiments

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Generation of pure GABAergic neurons by transcription factor programming.

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Immunofluorescence experiments were performed essentially as described5,10. Antibodies used include Calbindin (1:1,000, Swant, 300), Calretinin (mouse, 1:500, DAKO, M7245), Calretinin (mouse, 1:1,000, Swant, 6B3), Somatostatin (rabbit, 1:5,000, Bachem, T-4103.00), NKX2-1 (mouse, 1:300, Invitrogen, 18-0221), ISLET1 (mouse, 1:100, DSHB, 39.3F7), Pan DLX (rabbit, 1:1,000, gift from Y.M. Morozov, Department of Neurobiology, Yale University School of Medicine), NeuN (mouse, 1:100, Millipore, MAB377), GAD1 (mouse, 1:1,000, Millipore, MAB5406), GAD2 (mouse, 1:500, DSHB, GAD6), Synapsin-1 (rabbit, 1:1,000, Synaptic System, 106-002), DCX (rabbit, 1:1,000, Cell Signaling, 4604S), OLIG2 (rabbit, 1:1,000, Millipore, AB9610), TUJ1 (mouse, 1:1,000, Covance, MMS-435P), TUJ1 (rabbit, 1:1,000, Covance, MRB- 435P), VGAT (mouse, 1:1,000, Synaptic System, 131 003), Human nuclei antigen (mouse, 1:100, Millipore, MAB1281), GFP (chicken, 1:1,000, Aves Labs, GFP-1020), COUPTFII (mouse, 1:300, Perseus Proteomics, PP-H7147-00), MAP2 (chicken, 1:20,000, Abcam, AB5392), MAP2 (mouse, 1:500, Sigma Aldrich, M4403), GABA (rabbit, 1:1,000, Sigma Aldrich, A2052), Parvalbumin (mouse, 1:1,000, Sigma, P3088) (see also Supplementary Table 1).