RNA-sequencing of tamoxifen resistant LY2 cells transfected with siRNA-HOXC11.

To assess the global effects of HOXC11 in endocrine resistant breast cancer cells we performed RNA-seq on LY2 cells which were transfected with either siRNA targeting HOXC11 (siHOXC11) or a scrambled negative control siRNA (scrHOXC11) in the presence of 4-OH-tamoxifen (10-8M). Knockdown was verified by Taq-man qRT-PCR prior to library preparation. RNA (10µg) was extracted using an Oligotex mRNA kit (Qiagen) as per manufacturer’s instructions (n=4). RNA was reverse transcribed followed by mRNA library preparation and sequencing based on a protocol outlined by Wilhelm et al., 2010. Sequencing was performed on an Illumina Genome Analyzer II (GAII) (54 million reads per sample on average). Overall design: Silencing of HOXC11 in tamoxifen resistant LY2 cells to identify putative HOXC11 target genes.

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Ali A, Creevey L, Hao Y, McCartan D, O'Gaora P, Hill A, Young L, McIlroy M