Copy number abnormalities analysis of CD138⁺⁺ and CD138^low cells.

<p>Log ratio plot of all chromosomes corresponding to CD138⁺⁺ RPMI-8266 cells (n = 3; left panel) and CD138^low RPMI-8266 cells (n = 3; right panel) on the basis of Cytoscan HD array generated with Nexus.</p

Using Style to Understand Descriptions of Software Architecture

The software architecture of most systems is described informally and diagrammatically. In order for these descriptions to be meaningful at all, figures are understood by interpreting the boxes and lines in specific, conventionalized ways [5]. The imprecision of these interpretations has a number of limitations. In this paper we consider these conventionalized interpretations as architectural styles and provide a formal framework for their uniform definition. In addition to providing a template for precisely defining new architectural styles, this framework allows for the proof that the notational constraints on a style are sufficient to guarantee the meanings of all described systems and provides a unified semantic base through which different stylistic interpretations can be compared

Genes differentially expressed in CD138^low RPMI-8226 <i>versus</i> CD138⁺⁺ RPMI-8226 cells.

<p>Genes differentially expressed in CD138^low RPMI-8226 <i>versus</i> CD138⁺⁺ RPMI-8226 cells.</p

Proliferation of CD138⁺⁺ and CD138^low subpopulations.

<p>(A) Left: Cell percentage of CD138⁺⁺ and CD138^low RPMI-8226 or NCI-H929 cells in G0-G1, S and G2-M phases. The results are expressed as the means ± SEM of at least three independent experiments. Right: Representative DRAQ5 histograms for each indicated population. (B) Left: Relative Ki-67 MFI of CD138⁺⁺ and CD138^low RPMI-8226 and NCI-H929 cells with respect to isotype control. Results are expressed as the means ± SEM of three independent experiments. Right: Representative Ki-67 histograms for each indicated population. Filled histograms (isotype control); open histograms (Ki-67).</p

Tumorigenic potential of CD138⁺⁺ and CD138^low subpopulations.

<p>(A) 3×10⁴ sorted CD138⁺⁺ RPMI-8226 or CD138^low RPMI-8226 cells were subcutaneously injected into CB17-SCID mice to generate primary tumors. Subsequently, 3×10⁶ cells isolated from selected primary tumors were serially transplanted into new CB17-SCID mice to generate secondary tumors. The engraftment efficacy is indicated in each case. (B, C) Tumor growth curves for CB17-SCID mice that developed measurable primary and secondary tumors. Growth curves represent tumor volumes (means ± SEM; n = 4–6) until the time point in which the first mouse in every group is sacrificed.</p

Study of aldehyde dehydrogenase expression and drug sensitivity in CD138⁺⁺ and CD138^low subpopulations.

<p>(A) Single parameter histograms illustrating the expression of ALDH in the presence or absence of the ALDH inhibitor DEAB (diethylaminobenzaldehyde) in CD138⁺⁺ and CD138^low subpopulations of the cell lines RPMI-8226, NCI-H929 and MM1S (B) Sorted CD138⁺⁺ or CD138^low RPMI-8226 cells were incubated in the absence (control) or presence of bortezomib (10 nM), melphalan (10 μM) or doxorubicin (250 nM) for 24 and 48 hours. After the incubation time, cell viability was measured by MTT assay and the percentage of cell viability was calculated considering control as 100%. Results are the means ± SEM of at least three independent experiments.</p

CD138 expression in MM cell lines.

<p>(A) Dot plots showing the percentage of CD138⁺⁺ and CD138^low cells in MM cell lines. (B) Top: summary of the expression of CD19, CD20, CD27, CD38, CD45 and CD56 in CD138⁺⁺ and CD138^low subpopulations in MM cell lines. Code: – (negative); −/+ (heterogeneity); dim (weak positive); + (positive); ++ (high positive). Bottom: representative dot plots corresponding to non-stained RPMI-8226 cells (negative control; light grey) and stained CD138⁺⁺ (black) and CD138^low (dark grey) RPMI-8226 cells. (C) Expression of CD138 by real-time quantitative PCR in CD138⁺⁺ and CD138^low RPMI-8226 subpopulations. Relative values were calculated by the 2^−ΔCt method (ΔCt = Ct_(Gene)−Ct_(GAPDH)). The <i>GAPDH</i> gene was used as a control gene. Results are expressed as the means ± SEM (n = 3). (D–K) Confocal images corresponding to the immunocytochemistry for CD138 (red) in CD138⁺⁺ and CD138^low RPMI-8226 and NCI-H929 cells. Nuclear DNA was stained with DAPI (blue). Magnification of the lens, 63x. Specific “4× zoom” was made in E, G, I, K.</p