GD 12.5 mice were sacrificed and precursor of cortical neural epithelium was isolated and grown as Neurospheres
Growth protocol
We isolated cortical neuroepithelium from embryonic day 12.5 mouse fetuses (C57BL/6) from the dorsal telecephalic vesicles and cultured as neurosphere culture. Extreme care was taken to avoid structural precursors of hippocampus and striatum. Precursor cultures were established at an initial density of 106 cells in serum-free mitogenic media DMEM/F12 (catalog#11330-032; Invitrogen, Carlsbad, CA), 20 ng/ml bFGF (basic fibroblast growth factor; catalog #13256-029; Invitrogen), 20 ng/ml hEGF (human epidermal growth factor; catalog #53003-018; Invitrogen), 0.15 ng/ml LIF (leukemia inhibitory factor; catalog #L200; Alomone Labs, Jerusalem, Israel), ITS-X (insulin-transferrin-selenium-X) supplement (catalog #51500-056; Invitrogen), 0.85 Us/ml heparin (catalog #15077-019; Invitrogen), and 20 nM progesterone (catalog #P6149; Sigma, St. Louis, MO). Cells treated with antisense KO reagents for microRNA miR-335 and control for 24
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated from control and Treated samples Trizol method (#15596-026 Invitrogen)
Label
2 ug total RNA was used to generate biotin-labeled cRNA amplification protocol using the CodeLink iExpress Kit (Applied Microarray, Tempe, AZ).
Label protocol
Labeled cRNA was applied to CodeLink (Mouse) genome arrays for an 18 h hybridization followed by washing, staining, and scanning as per the CodeLink protocol.
Hybridization protocol
CodeLink protocol
Scan protocol
CodeLink protocol
Description
Array images were processed using CodeLink system software and global median normalization was used to generate normalized expression values
Data processing
Array images were processed using CodeLink system software and global median normalization was used to generate normalized expression values
