Low input quickamp one color labelling kit

Microarrays analysis were performed from RNAs extracted from cells using STAT 60 and RNeasy Mini Kit (50) from QIAGEN and following the manufacturer’s instructions protocol. 200 ng of total RNA was labelled using Agilent low input QuickAmp one-color labelling kit. Quality of labelling checked using nanodrop 8000 and hybridised onto an Agilent Human GE 4x44K v2 microarrays at 65 °C for 16 h. Slides were washed and scanned using a GenePix pro- 4100A scanner, quality controls and raw data of the scanned images were generated using Agilent feature extraction software V11.0. Raw data was normalised using Partek Genomic Suit V 6.0 and used for ANN analysis. Microarrays gene expression data were used for pathways analysis using Panther Pathway. Gene expression of IRF-9, PTK2B, C2ORF42 and DHFR2 were assessed by Real-time quantitative PCR using the following primers: FH1_IRF9:5′-CTCAGAAAGTACCATCAAAGC-3′; RH1_IRF9:5′- TCATTATTGAGGGAGTCCTG-3′; FH1_PYK2: 5′-AATGCACTTGACAAGAAGTC-3′; RH1_PYK2:5′-GCTTTAAGTTCTCCTGCATC-3′; FH1_C2orf42: 5′-AGCTTTTGTTCGGAAAGATG-3′; RH1_C2orf42: 5′-GCATCTCTGGGGTATCTAAG-3′; FH1_DHFR2:5′-CGCTGTGTCCCAAAACATGG-3′; RH1_DHFR2: 5′- GAATTCATTCCTGAGCGGCG-3′. The gene expression microarrays data was deposited in the online database ArrayExpress under the accession number: E-MTAB-7737.