Prior to MoFlo sorting, single cell suspensions were stained with the following antibodies: FITC-conjugated anti-CD56 (HCD56) (Biolegend, San Diego, USA), PE-conjugated anti-CD16 (368) (Biolegend, San Diego, USA) APC-conjugated anti-CD3 (UCHT1) (Biolegend, San Diego, USA) PE-Cy7-conjugated anti-CD19 (HIB19) (Biolegend, San Diego, USA). NK cell purity, functional properties and subset distribution was analyzed by FACS Canto II (BD Biosciences, Heidelberg, Germany) and use of the FlowJo Software (Treestar, Ashland, Oregon, USA).
Extracted molecule
total RNA
Extraction protocol
RNA was isolated from freshly sorted NK subsets using the RNeasy Mini/Micro Kit (QIAGEN, Hilden, Germany) according to the manufacturer's instructions. RNA quality was checked using the Agilent 2100 Bioanalyzer.
Label
biotin
Label protocol
3' IVT Express.
Hybridization protocol
GeneChip Hybridization, Wash and Stain.
Scan protocol
GeneChip Scan 3000 7G.
Description
Gene expression of freshly sorted NK cells.
Data processing
RMA normalization algorithm was applied through the affy package from R. The limma package from R was used for statistical calculations.
