Additional Supporting Information may be found in the online version of this article.

HEP_24466_sm_SuppInfoFigS1.tif1271KFigure S1. Hazard functions for death and metastasis with Weibull model. The estimated hazard function for death (A) and metastasis (B) are shown. For death, the hazard function was increased with a rate decreasing over time (A), whereas for metastasis, the hazard is very high at the beginning but it rapidly decreases over time (B).
HEP_24466_sm_SuppInfoFigS2.tif4355KFigure S2. Immunocytochemical expression of K19 in human CCA cell lines is not altered by lentiviral silencing of S100A4. K19 expression (green) was similar in EGI-1 (A), sh8-EGI-1 (B) and sh9-EGI-1 (C) cells as well as in TFK-1 (D) cell line. Nuclei were counterstained with DAPI (blue). Original magnification: 200×.
HEP_24466_sm_SuppInfoFigS3.tif625KFigure S3. Functional effects of S100A4 silencing in EGI-1 cells as compared to TFK-1 cells on cell proliferation. Using MTS assay after a 24h incubation with growth medium, no difference in the proliferative activity was found among silenced, scramble, EGI-1 and TFK-1 cells.
HEP_24466_sm_SuppInfoFigS4.tif2193KFigure S4. MMP-2 and MMP-9 expression in EGI-1 cells and functional effects of S100A4 silencing in EGI-1 cells on MMP-9 secretion. Tissue sections of liver metastases derived from xenotransplantation of EGI-1 cells in SCID mice immunostained for MMP-2 (A) and MMP-9 (B) showed that MMP-9, but MMP-2, was strongly expressed by metastasizing CCA cells. Immunoperoxidase technique, original magnification: 200×. In the supernatant of cultured cells (n×4), MMP-9 secretion by EGI-1 cells assessed by ELISA was significantly reduced after lenitiviral silencing of S100A4, as observed in sh8 and sh9 clones (p<0.001); in TFK-1 cells, MMP-9 secretion is negligible (C).
HEP_24466_sm_SuppInfoTableS1.doc39KTable S1. Summary of survival for each group in Kaplan-Meier survival curves.
HEP_24466_sm_SuppInfoTableS2.doc72KTable S2. Clinical features of the CCA series (n=67) according to S100A4 grouping considered for NPMLE analysis and Weibull model a.
HEP_24466_sm_SuppInfoTableS3.doc51KTable S3. Results of Weibull Model with outcome as death, in 67 resected human CCA.
HEP_24466_sm_SuppInfo.doc27KSupporting Information

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