Total RNA was extracted using the RNeasy Plus Mini Kit (QIAGEN) and treated with a TURBO DNA-free Kit (Ambion). cDNA was synthesised from RNA using the PrimeScript RT Reagent Kit (TaKaRa). Quantitative PCR (Q-PCR) was performed with SYBR Green (Applied Biosystems) using an ABI 7700 Prism real-time PCR instrument. The expression of mRNA was normalised to the expression of either 18 S ribosomal RNA or GAPDH as the change in cycling threshold (ΔCT) method and calculated based on 2-ΔCT. Results are articulated as relative gene expression for triplicates of each target. PCR primers used are listed in Supplementary Table S4.
