Figure S1. (A) RNA interference (RNAi) treatment [using double-stranded RNA (dsRNA) probe of hormone receptor-like in 96 (hr96)] of S2 cells in conjunction with the phenobarbital (PB) promoter assay. Drosophila S2 cells were cultured in a 25 cm2 flask with Cellfectin ± dsRNA probe. Cells were subcultured every 3 days and Cellfectin ± dsRNA probe were added. RNAi-treated cells and control cells of 3, 6, 9 and 12 days were subjected to PB promoter assays using the promoter construct −330/+85. Transfected RNAi-treated cells were continued with treatment of dsRNA probe. After 48 h incubation with or without PB, firefly and Renilla luciferase activities were measured. (B) PB induced promoter activities (grey bars) and basal promoter activities (white bars) derived from the use of control cells and RNAi-treated cells of 3-, 6-, 9- and 12-day RNAi treatments (using dsRNA probe of hr96). Bars represent mean of promoter activity [firefly luminescence units (LU)/Renilla LU] ± SD of three independent transfections (n = 3). These results show that promoter assays using 3-, 6-, 9- and 12-day RNAi-treated cells resulted in 1.5, 10.5, 19.4 and 25.4% reduction of PB induction, respectively, compared to control cells.

IMB_1047_sm_Figure_S1a.TIF109KSupporting info item
IMB_1047_sm_Figure_S1b.TIF105KSupporting info item

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