<i>Let-7a</i> real-time PCR analyses.

<p>Relative <i>let-7a</i>/RNU6B expression in native CT1258, CT1258-EGFP and CT1258-HMGA2-EGFP cells. Error bars are standard deviations. No statistical significant expression deregulation of <i>let-7a</i> in CT1258-EGFP and CT1258-HMGA2-EGFP was detected when compared to native CT1258 cells. Statistical significant p value was defined as ≤0.05.</p

<i>HMGA1</i> real-time PCR analyses.

<p>Relative <i>HMGA1/HPRT1</i> and <i>HMGA1/GUSB</i> expression in native CT1258, CT1258-EGFP and CT1258-HMGA2-EGFP cells. Error bars are standard deviations. *p≤0.05 indicates a statistical significant increased expression of <i>HMGA1</i> in CT1258-HMGA2-EGFP cells when compared to native CT1258 and CT1258-EGFP.</p

Cytogenetic analyses.

<p>Metaphase spreads derived from CT1258 (A), CT1258-EGFP (B) and CT1258-EGFP-HMGA2 (C) cells after GTG-banding. The arrows indicate the centromeric fusions between the canine chromosomes 1 and 5 (der(1;5)) and a large bi-armed marker (mar) consisting of material from chromosomes 1 and 2 being characteristic for the CT1258 cell line.</p

<i>HMGA2</i> real-time PCR analyses.

<p>Relative <i>HMGA2/HPRT1</i> and <i>HMGA2/GUSB</i> expression in native CT1258, CT1258-EGFP and CT1258-HMGA2-EGFP cells. Error bars are standard deviations. *p≤0.05 indicates a statistical significant expression deregulation of <i>HMGA2</i> in CT1258-HMGA2-EGFP cells when compared to native CT1258.</p

BrdU cell proliferation assay.

<p>Measured proliferation of native CT1258, CT1258-EGFP and CT1258-HMGA2-EGFP cells. A statistical significant increased proliferation was detected for CT1258 cells expressing the EGFP-HMGA2 fusion protein in comparison to native CT1258 and EGFP expressing CT1258 cells. Each bar represents a mean ± SD, *p≤0.05, ***p≤0.001.</p

Details of CT1258 chromosomal aberrations.

<p>Detailed presentation of chromosomal aberrations found in metaphases of CT1258, CT1258-EGFP and CT1258-EGFP-HMGA2. Two derivative chromosomes (der(1;5)) and the large bi-armed marker chromosome (mar) were found in each cell line.</p

Immunocytochemical staining.

<p>A: Native CT1258 cells, B: CT1258-EGFP cells, C: CT1258-EGFP-HMGA2 cells. Approximately 50% of the native CT1258 cell line and of CT1258-EGFP cells showed a HMGA2-positive nuclear labelling. In approximately 70–80% of CT1258-EGFP-HMGA2 cells, a strong and exclusively nuclear labelling for HMGA2 was detectable.</p