19 research outputs found
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>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
<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
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
<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
PCR for antigen receptor rearrangement. PARR of CLBL-1 cells (A), Xenograft-1 (tumor of right flank,B), Xenograft-2 (tumor of right flank,C), CLBL-1M cells (D) and the OSW cell line (E).
<p>Lane 1 shows Cµ serving as positive control for DNA. Lanes 2 and 3 show bands of IgH and TCRγ PCR products, respectively.</p
BrdU cell proliferation assay. Measured proliferation of CLBL-1 (A, B, C) and CLBL-1M (D, E, F) after stimulation with DSP30 and/or IL-2 at different time points (48 h, 72 h, 96 h) in comparison to untreated cells. Each bar represents a mean ± SD, *p≤0.05, **p≤0.001 to 0.01.
<p>BrdU cell proliferation assay. Measured proliferation of CLBL-1 (A, B, C) and CLBL-1M (D, E, F) after stimulation with DSP30 and/or IL-2 at different time points (48 h, 72 h, 96 h) in comparison to untreated cells. Each bar represents a mean ± SD, *p≤0.05, **p≤0.001 to 0.01.</p
Monoclonal antibodies used for flow cytometry for CLBL-1 before inoculation of the cells, after sacrificing the inoculated mice and after cultivating the isolated cells <i>in vitro</i> (CLBL-1M).
*<p>Fluorescence labelling was achieved by use of a secondary antibody.</p><p>Abbreviations: m  =  mouse; r  =  rat; FITC  =  Fluorescein isothiocyanate, APC  =  Allophycocyanin;</p><p>PE  =  Phycoerythrin.</p>**<p>cross-reactivity pattern for canine lymphocytes lymphocytes <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040078#pone.0040078-Saalmller1" target="_blank">[36]</a>.</p