For cell_M5A, M5K, M6C, M6H, pCAG-NICD and pEF-p18 was co-electroporated into dorso-lateral part of cerebral wall at E11 by in vivo electroporation. Dorso-lateral portion of cerebral wall of CD1 mice at E11,E12, small VZ/SVZ fragments of the same portion at E14 (3 days after electroporation), E16, or 3 days-in-vitro neurospheres were digested in 100ul of 0.25%Trypsin/0.5% Glucose /PBS for 5min at 37°C and triturated. After adding Hanks solution (Nakarai, Japan) with trypsin inhibitor (Ovomucoid, Sigma), singly-isolated cells were randomly picked up manually by glass-capillary under inverted microscope and transferred into PCR tubes containing 4.5ul of cell lysis buffer.
Growth protocol
For cell_W2G,W2O and W6D, cells from dorso-lateral portion of cerebral wall of E11 CD1 mice were cultured for 3 days in DMEM/F12 with N2, B27, 1mM N-acetylcysteine, 20ng/ml FGF2 and 20ng/ml EGF in suspension culture dish.
Extracted molecule
total RNA
Extraction protocol
Single cell was lysed in a PCR tube containing 4.5ul of cell lysis buffer containing spike RNA: poly(A)-tailed Bacillus subtilis lys, dap, phe, and thr RNAs at 1000,100, 20, and 5 copies per cell, respectively, and the clude lysate was used for cDNA synthesis and amplification as descibed (Kurimoto et al.Nucreic Acid Res.34:e42, 2006).
Label
biotin
Label protocol
cDNA samples were subjected to the one-cycle target labeling procedure for biotin labeling by in vitro transcription (Affymetrix)
Hybridization protocol
RNA was subsequently fragmented and hybridized to the GeneChip according to manufacturer’s instructions. GeneChips were washed and stained in the Affymetrix Fluidics Station 450
Scan protocol
The microarray image data were processed with the GeneChip Scanner 3000 (Affymetrix).
Description
Gene expression data of single apical progenitor (AP) cell derived from dorso-lateral portion of cerebral wall of E12 CD1 mouse
Data processing
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings.
