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FilenameFormatSizeDescription
HEP_25933_sm_SuppFig1.tif5150KSupporting Information Figure 1. (A) Increased SOX1 expression dramatically suppressed anchorage-independent growth (AIG) of HepG2, Huh7 and SK-Hep-1 cells. The representative photographs were taken after 4 weeks of incubation and the number of colonies was measured. (B) Elevated SOX1 expression inhibited the invasion ability of SK-Hep-1 and HA22T cells. A representative photograph of invasive cells on the lower surface of the insert is shown.
HEP_25933_sm_SuppFig2.tif649KSupporting Information Figure 2. Tumors harvested on day 38 after implantation were used to verify the level of SOX1 expression by Western blot. β-actin was used as a loading control.
HEP_25933_sm_SuppFig3.tif463KSupporting Information Figure 3. Time- and dose-dependent induction of SOX1 protein expression by doxycycline. (A) DOX (0, 0.01, 0.1,1 and 10 μg/ml) was applied to cells for 48 hr. (B) Hep3B cells were treated with DOX (1 μg/ml) for 0, 12, 24, 48 and 72 h. Cell lysates were subjected to Western blot analysis. β-actin was used as a loading control.
HEP_25933_sm_SuppFig4.tif2609KSupporting Information Figure 4. (A) Inducible SOX1 expression dramatically suppressed anchorage-independent growth of HepG2, Huh7 and SK-Hep-1 cells. The representative photographs were taken after 4 weeks of incubation and the numbers of colonies were measured. (B) A Matrigel invasion assay was performed in SK-Hep-1 cells inductively expressing SOX1 by treatment with doxycycline 1 μg/ml for 3 days. The cells were then placed in a Matrigel-coated Boyden chamber and allowed to invade for 24 hr. A representative photograph of invasive cells on the lower surface of the insert is shown.
HEP_25933_sm_SuppFig5.tif1346KSupporting Information Figure 5. (A) The SOX1 protein levels expressed inductively by 1 μg/ml DOX for 7 days (DOX+) and then withdrawal of DOX for a further 7 days (DOX withdrawal) were investigated by Western blot. (B) A representative photograph of the AIG assay from cells treated as described in (A) is shown.
HEP_25933_sm_SuppFig6.tif1236KSupporting Information Figure 6. GST pull-down assay. Purified GST only or GST-SOX1 proteins were immobilized on a glutathione-sepharose column and mixed with lysate from Huh6 cells. After washing and elution, the remaining proteins were analyzed by Western blot using anti-β-catenin and anti-GST antibodies.
HEP_25933_sm_SuppFig7.tif1588KSupporting Information Figure 7. Truncated Flag-SOX1 suppresses ß-catenin-mediated TCF/LEF signaling. (A) Full-length Flag-SOX1 and truncated Flag-SOX1 as indicated were established and shown by Western blot from COS7 cells. (B) Huh6, Hep3B and HepG2 cells were transfected with Flag-tagged plasmids as indicated and the TOPFLASH reporter gene and dual-luciferase activities were measured as described above. The data showed that full-length Flag-SOX1, Flag-?N-SOX1 and Flag-?C-SOX1 could suppress ß-catenin-mediated TCF/LEF signaling with an efficiency comparable with that of Flag-Vector, whereas Flag-C-SOX1 could not suppress it. The luciferase activity was normalized to the Renilla luciferase activity. The results are presented as the mean ± SE. Experiments were performed in triplicate. Significant differences are indicated by asterisks; (*) for P < 0.05 and (*) for P < 0.01.
HEP_25933_sm_SuppTab1.tif6461KSupporting Information Table 1.
HEP_25933_sm_SuppTab2.doc69KSupporting Information Table 2. Correlation between SOX1 expression and clinicopathologic characteristics
HEP_25933_sm_SuppInfo.doc70KSupporting Information

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