Gal-3 knockdown decreases anchorage independent cell growth, but has no effect on tumor growth in a xenograft mouse model.

<p>(<b>A</b>) Gal-3 was transiently silenced in pancreatic cancer cell lines using two independent siRNA sequences (siGal-3 1 and 2). Knockdown and control cells were seeded in soft agar and colony formation assessed after 7 days. Results were normalized to control siRNA transfected cells (NC). Data includes a minimum of three independent experiments. * Indicates p<0.05 as compared with control siRNA transfected cells (double-sided unpaired <i>t</i>-test). (<b>B</b>) Western blot analysis of nuclear (n) and cytoplasmatic (c) protein fractions derived from pancreatic cancer cell lines. Gal-3 is shown in the upper lane. β-actin staining was used as loading control. Staining for the nuclear factor ORC2 was used to assess the purity of the subcellular fractions. (<b>C</b>) Xenograft tumors were induced in nude mice by subcutaneous injection of two stable Gal-3 knockdown S2-007 cell clones (shGal-3 2 and 3) and two nonsilencing control shRNA transfected S2-007 clones (shC 2 and 3). Six mice per experimental group were analyzed. Box-and-whisker plots illustrate tumor volumes on day 22 post injection. Whiskers denote 1.5× interquartile range (IQR). Values outside this range are shown as filled triangles. (<b>D</b>) Gal-3 mRNA levels in bulk tissue of xenograft tumors where determined by qRT-PCR. RPLP0 was used as the reference gene.