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HEP_23443_sm_Suppfig1.tif602KSupplementary Fig. S1 Expression of mature miR-30d in various HCC cell lines The relative expression level of mature miR-30d in various liver cancer cell lines was determined by quantitative Real-time PCR. Data are normalized by U6 snRNA respectively.
HEP_23443_sm_Suppfig2.tif2285KSupplementary Fig. S2 Establishment of stable cell lines highly expressing miR-30d A. The relative expression level of mature miR-30d in Huh-7, SMMC-7721 or SK-HEP-1 cells infected with lentivirus expressing either miR-30d or mock vector. B. The mature miR-30d expression level in stable SMMC-7721 cells under the pressure of G418. Quantitative RT-PCR assay is used, data are normalized by U6 snRNA, and fold changes are indicated in the form of mean ± S.E.M.
HEP_23443_sm_Suppfig3.tif756KSupplementary Fig. S3 Over-expression of miR-30d has no obvious effects on HCC cell growth in vitro A. The CCK-8 assay of SK-HEP-1 cells infected with lentivirus expressing miR-30d or mock control. One thousand cells per well are seeded, and measured every day. Mean values are plotted as shown, and the bars indicate S.E.M in triplet. B. The colony formation assay of SMMC-7721 cells infected with lentivirus expressing miR-30d or mock control. Five hundred cells per well are seeded and cultivated for 2 weeks. After fixation and dying, colonies of miR-30d are counted and normalized by mock control. The bars represent S.E.M from three independent experiments.
HEP_23443_sm_Suppfig4.tif3101KSupplementary Fig. S4 Sixty-nine genes that were down-regulated by miR-30d in HCC stable cells A. The expression profiles of 69 down-regulated genes in two stable clones of SMMC-7721 cells expressing miR-30d. Green color represents the ratio of these down-regulated genes in SMMC-7721-30d/SMMC-7721-Mock. Sharper the color, lower the ratio. B. The name and expression information of these 69 genes are listed.
HEP_23443_sm_Suppfig5.tif1209KSupplementary Fig. S5 Expression of GNAI2 in various HCC cell lines The relative expression level of GNAI2 in various HCC cell lines was determined by quantitative Real-time PCR. Data are normalized by β-actin respectively, and the bars stand for S.E.M.
HEP_23443_sm_Suppfig6.tif785KSupplementary Fig. S6 GNAI2 interaction with miR-30 family A. Schema of predicted binding sites of miR-30 family within the 3′UTR of GNAI2 by three online prediction algorithms TargetScan, PicTar and MicroCosm Targets. B. Western blot assays of endogenous GNAI2 protein level in SK-HEP-1 cells after transfected with miR-30a, b, c, e or negative control.
HEP_23443_sm_Supptext.doc113KSUPPLEMENTARY MATERIALS

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