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. 2011 Jun 16;6(6):e20859. doi: 10.1371/journal.pone.0020859

Figure 7. Gal-3 knockdown decreases anchorage independent cell growth, but has no effect on tumor growth in a xenograft mouse model.

Figure 7

(A) 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 t-test). (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. (C) 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. (D) Gal-3 mRNA levels in bulk tissue of xenograft tumors where determined by qRT-PCR. RPLP0 was used as the reference gene.