Validation of gene expression by quantitative real-time PCR, Western blotting, and flow cytometry. To validate gene expression data, quantitative real-time PCR for aurora-A, -B, -C in (Ai) 10 human myeloma cell lines (HMCLs) and (Aii) 10 primary multiple myeloma cell samples (pMMCs) was performed. Shown are -dCt values (reference gene 18S-RNA). (B) Gene expression data were further validated by Western blotting. Shown are the blots of 10 cell lines for aurora-A and -B with β-actin as a loading control and HELA cells as a positive control. (C) Intracytoplasmatic expression of aurora-A and -B as determined by flow cytometry. Shown is the cell line OPM-2. The light gray line indicates control without primary antibody; the black line, measurement with primary and secondary antibody.