To explore TIMELESS's potential functional significance in regulating cancer-relevant gene networks, we performed a loss-of-function analysis using TIMELESS-targeting siRNA oligos, followed by a whole genome expression microarray and subsequent network analysis.
A whole genome expression microarray (Agilent, Inc 44k chip, performed by MoGene, LC, St Louis, MO) was used to interrogate gene expression in cells with normal and reduced CRY2.
The Illumina whole genome expression microarray platform was used to assess differential expression.
A more quantitative assessment of genome-wide differential gene expression will be facilitated by interrogations of whole genome expression microarrays, which are in progress (W. Miao, M. Gorovsky, et al.; pers. comm).
A loss-of-function analysis was then performed using TIMELESS-targeting siRNA oligos followed by a whole-genome expression microarray and network analysis.
To determine TIMELESS's role in tumorigenesis, we then performed an in vitro loss-of-function analysis using TIMELESS-targeting siRNA oligos followed by a whole-genome expression microarray.
To elucidate the comprehensive regulation in transcriptional profile induced by AGO2, whole-genome expression microarray analysis was conducted between the CNE2Z cells transfected with AGO2 shRNAs and control shRNA.
In the current study, we report our findings from the expression profiling analysis of TIMELESS in different tumor types using publically available online tools and microarray datasets, and a loss-of-function analysis using TIMELESS-targeting siRNA oligos followed by a whole-genome expression microarray and network analysis.
Validation of whole-genome expression microarray was performed by qRT-PCR on a representative list of significant genes.
The application of whole-genome expression microarray usually produces large data sets, which are very difficult to analyze manually.
