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. 2008 Sep 16;13(11-12):4608–4622. doi: 10.1111/j.1582-4934.2008.00499.x

Figure 7.

Figure 7

mRNA expression levels of cell cycle-related genes in MSP58 depleted U251 cells. (A) cDNA array analysis demonstrated differentially expressed cell cycle-related genes in MSP58 depleted U251 cells. Total RNA (2 μg) from U251-NC or U251-S cells was reverse transcribed to cDNA in the presence of Biotin-16-UTP. Labeled cDNA was hybridized to cell cycle-specific gene expression array membranes. After normalization of the mRNA levels of a given gene to the average value of an internal control on the same membrane, gene-expression levels in MSP58-depleted cells were calculated in arbitrary units divided by the values from control cells. A two-fold change of gene expression comparing U251-S to U251-NC cells was considered significant. (B) Microarray results were validated by real-time RT-PCR. Total RNA was extracted from U251-NC or U251-S glioma cells, and real-time quantitative RT-PCR was performed to confirm the change of different cell cycle-related gene obtained by cDNA array analysis. mRNA expression of various genes was normalized to β-actin mRNA expression. Results were expressed as mean ± SD from three independent experiments, each performed in duplicate.