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. Author manuscript; available in PMC: 2009 Jun 8.
Published in final edited form as: Oncogene. 2009 Jan 26;28(12):1561–1568. doi: 10.1038/onc.2008.497

Figure 1. Downregulation of Singleminded-2s (SIM2s) expression in HRAS-transformed MCF10A cells.

Figure 1

(a) Altered morphology of Harvey-Ras (Ha-Ras)-transformed MCF10A cells. Cells were transduced with either empty vector (pLPCX, left panel) or a vector containing the Ha-Ras coding sequence (HRAS, right panel). Differential contrast imaging of cells following 2 days of selection in medium containing 1 µg/ml puromycin, cells, showed that Ha-Ras-transduced cells lost their normal ovoid shape and displayed abnormal cellular organization. Ha-Ras construct was provided by Brian and Alana Welm and subcloned into EcoRI sites in pLPCX (Clontech). (b, c) SIM2s and E-cadherin expression is decreased in Ha-Ras-overexpressing MCF10A cells. Total RNA and protein were isolated from control (pLPCX) and Ha-Ras-overexpressing cells (HRAS) and analysed for SIM2s expression by quantitative real-time RT-PCR and western analysis (Chemicon) as previously described (Laffin et al., 2008). The results show that SIM2s and E-cadherin expression are significantly reduced in Ha-Ras-transduced MCF10A cells in comparison to control cells (mean ± s.e. for three replicates; *P=0.0002).