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Immunocytochemistry and imaging Hypothalamic progenitors were re-plated onto PhenoPlate TM 96-well microplates (Perkin Elmer) and hypothalamic cultures at 33-40 days post-differentiation were fixed in 4% paraformaldehyde (Thermo Fisher Scientific) for 10 minutes, rinsed three times with Tris-buffered saline containing 0.1% Triton X-100 (TBS-T

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[0041] The solvent used in the recrystallization of step 3) is chosen from Et 2 O, CH 2 Cl 2 , CHCl 3 , MeOH, CH 3 CN, THF, EtOAc, EtOH, Toluene, Hexane, Acetone or a mixed solvent of any thereof, preferably a mixed solvent of MeOH and CH 2 Cl 2