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Fig. S5 Bcdo2 mRNA expression in T47D cells. (A) T47D cells were incubated in the presence of vehicle only (0.1% v/v DMSO and 0.1% v/v THF), 1 μM of either β,β-carotene or canthaxanthin for 2 hours at 37°C and 5% CO2. (B) Alternatively, T47D cells were pre-treated with siRNA specific to human BCDO2 for 72 hours as outlined in the Materials and methods. Cells were then incubated in the presence of vehicle or carotenoids as described in A. Cells were harvested in 1×PBS and total RNA was isolated. Bcdo2 mRNA expression was evaluated by quantitative real-time RT-PCR using a Bcdo2 gene-specific probe set. The 18s rRNA probe set was used as the endogenous control. Post PCR products were electrophoresed on 2% (w/v) agarose gels and visualized by ethidium bromide staining. *P<0.05 compared with controls.