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Figure 2 | BMC Molecular Biology

Figure 2

From: SLUG: a new target of lymphoid enhancer factor-1 in human osteoblasts

Figure 2

Effect of Lef-1 overexpression on SLUG expression in hOBs. The effect of Lef-1 overexpression was examined at mRNA (A) and protein (B) level. (A) SLUG mRNA was evaluated by quantitative RT-PCR in hOBs and SaOS-2 osteoblast-like cells transfected with 2.5 μg of hLef-1 (K14-myc-hLEF1) expression plasmid. The cDNA obtained from total RNA was subjected to quantitative TaqMan RT-PCR for SLUG transcript analysis. The expression levels were normalized on the basis of GAPDH expression and results of the experiments are reported as relative mRNA expression levels. Results are representative of three independent experiments carried out in triplicate. ΔΔCt method was used to compare gene expression data; standard error of the mean (SEM) was calculated. * = p < 0.05. (B) SLUG protein levels were examined by Western blot analysis in hOBs and SaOS-2 osteoblast-like cells transfected with 2.5 μg of hLef-1 expression plasmid. Whole cell lysates were prepared and 25 μg of protein run on a 12% SDS-polyacrylamide gel. The proteins were visualized using Supersignal West Femto Substrate (Pierce). The quantitative presentation of the protein levels were performed by densitometric analysis using Anti-IP(3)K as control. D.U. = densitometric units. This experiment was repeated three times with similar results. A representative SLUG and Lef-1 Western blot analysis with size markers (KDa) is reported. * = p < 0.05.

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