cell line: HeLa cell type: cancer stem cells transfection: ZsGreen-cODCdegron
Growth protocol
HeLa cells transfected with ZsGreen-cODCdegron were cultured in DMEM supplemented with 10% FBS and 1% P/S.
Extracted molecule
total RNA
Extraction protocol
ISOGEN was used for extracting total RNA
Label
Cy5
Label protocol
Extracted total RNA was labeled with Cy5 using the 3D-Gene miRNA labeling kit (Toray, Kamakura, Japan)
Hybridization protocol
Hybridized for 16 h at 32 C with rotary shake (250 rpm). Hybridization buffer and washing protocol was followed by the protocol supplied by TORAY Industries, Inc..
Scan protocol
3D-Gene Scanner ((Toray Industries Inc., Tokyo, Japan) was used for scanning. Images were quantified using Extraction(Toray Industries Inc., Tokyo, Japan).
Data processing
The raw data of each spot was normalized by substitution with a mean intensity of the background signal determined by all blank spots’ signal intensities of 95% confidence intervals.
