{miR-124 RNA duplex (Pre-miR) was transfected into HepG2 cell line. The Pre-miR RNA duplex mimics the miR-124 precursor.miR-124 is highly expressed in brain and kidney (34), and it does not express in the cell line we studied. miR-124 overexpression profiles were examined at 4, 8, 16, 24, 32, 72 and 120 h post transfections. A negative control RNA duplex was also transfected as reference for each of these time points.Expression profiles were first examined at 72 h post transfection, and hundreds of genes were shown to be downregulated by miR-124 overexpression.We also performed real-time RT-PCR to validate all predicted targets downregulated at early stages of transfections. Figure 2 is a summary of the real-time RT-PCR validation results. The expression levels at 24 and 48 h were presented as percentages of the expression levels at 4 h. Among the validated targets, the microarray data indicated that genes 8801 and 8992 were first downregulated at 8 h; genes 8763, 3915, 6566, 9341, 10 449 and 10 634 were first downregulated at 16 h; genes 6836, 60 481 and 84 061 were first downregulated at 24 h; genes 27 230 and 55 225 were first downregulated at 32 h (underexpression is defined as at least 50% reduction of gene expression by array measurement).Thus, the real-time RT-PCR results agreed well with the microarray results.}{We also performed real-time RT-PCR to validate all predicted targets downregulated at early stages of transfections. Figure 2 is a summary of the real-time RT-PCR validation results. The expression levels at 24 and 48 h were presented as percentages of the expression levels at 4 h. Among the validated targets, the microarray data indicated that genes 8801 and 8992 were first downregulated at 8 h; genes 8763, 3915, 6566, 9341, 10 449 and 10 634 were first downregulated at 16 h; genes 6836, 60 481 and 84 061 were first downregulated at 24 h; genes 27 230 and 55 225 were first downregulated at 32 h (underexpression is defined as at least 50% reduction of gene expression by array measurement).Thus, the real-time RT-PCR results agreed well with the microarray results.}