Table 2 Partial list of miRNAs with significantly different CDK activity levels detected in SP of HCC cells compared to fetal liver cells microRNA SAM score Fold change False discovery rate (FDR) % hsa-miR-935 0.66 4.32 0.51 mmu-miR-10b 1.00 3.88 0.07

mmu-miR-21 0.80 2.96 0.00 mmu-miR-470* 0.69 2.81 0.00 hsa-miR-34c-3p 0.78 2.79 0.00 hsa-miR-650 0.76 2.71 0.00 hsa-miR-92b* 0.69 2.65 0.03 hsa-miR-193b 0.71 2.59 0.00 hsa-miR-374a* 0.68 2.58 0.24 hsa-miR-548c-3p 0.70 2.54 0.00 hsa-miR-33b 0.66 2.53 0.57 mmu-miR-199a-3p 0.71 2.52 0.00 hsa-miR-330-3p 0.71 2.51 0.00 mmu-miR-376a 0.69 2.48 0.13 mmu-miR-100 0.68 2.44 0.16 mmu-miR-717 0.66 2.36 0.62 mmu-miR-125b-5p check details 0.66 2.35 0.45 mmu-miR-449a 0.64 2.35 1.09 hsa-miR-21* 0.63 2.31 1.29 mmu-miR-883b-3p 0.63 2.29 1.20

mmu-miR-31 0.59 2.25 2.45 mmu-miR-34b-3p 0.57 2.14 3.43 mmu-let-7i* 0.55 2.02 4.66 hsa-miR-549 -0.70 0.05 2.84 mmu-miR-207 -0.86 0.23 6.02 mmu-miR-200a* -0.94 0.29 1.22 mmu-miR-207 -0.86 0.23 0.60 hsa-miR-148b* -0.76 0.36 2.72 mmu-miR-135a* -0.69 0.38 2.92 Figure 2 SAM outputs. SAM plotsheet outputs under the four sets of criteria: Δ = 0.25, fold change = 2. Conditions are indicated at the upper right corner of each plotsheet. The red, green, and black dots represent upregulated, downregulated, and insignificantly changed miRNAs, respectively. The upper and lower 45° degree lines indicate the Δ threshold R406 datasheet boundaries. The number of significant miRNAs, median number of false positives, and false discovery rate (FDR) are indicated at the upper left corner of the plotsheet. Figure 3 Heat map of altered miRNA expression. A heat map was generated using the expression ratios of 78 miRNAs Cyclooxygenase (COX) that differed significantly in SP of HCC cells compared to fetal liver cells, according to significance analysis of microarrays

(SAM). Red, overexpressed miRNAs; green, underexpressed miRNAs compared to counterparts. Relatedness in miRNA expression across samples is shown by a hierarchical tree on the Y axis through standard linkage. Validation of the differentially expressed miRNAs by qRT-PCR Using a stringent cut-off of P < 0.05, we found significantly altered expression of only 7 of all rat miRNAs analyzed in SP of HCC cells. In detail, five miRNAs were significantly up-regulated (miR-21, miR-34c-3p, miR-470*, miR-10b, let-7i*) and two miRNAs significantly down-regulated in SP of HCC cells (miR-200a*, miR-148b*). miRNA-specific qRT-PCR was used to validate the significantly altered miRNAs from the miRNA microarray results.