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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 2. Knockdown of Singleminded-2s (SIM2s) in MCF10A mammary epithelial cells by short-hairpin RNA (shRNA) disrupts three-dimensional acinar morphogenesis and induces a partial epithelial-mesenchymal transition (EMT).

Figure 2

(a, b) Inhibition of SIM2s expression in MCF10A cells by shRNA. MCF10A cells transduced with a confirmed SIM2-specific construct (SIM2s-sequence 3116; SIM2i) showed a significant reduction in SIM2s mRNA and protein as compared to a nonspecific control shRNA retroviral construct (Scr) (Laffin et al., 2008). (c) Low- and high-power differential interference contrast images of MCF10A acini grown on matrigel demonstrate that acinar morphogenesis is disrupted in SIM2i MCF10A cells as compared to Scr control. (d) SIM2i MCF10A cells display an increased ability to invade through a matrigel matrix using transwell invasion assays (Kwak et al., 2007). (e) Loss of SIM2s induces a partial EMT. Western blot analysis of Scr and SIM2i cells found that E-cadherin expression is reduced and vimentin expression is increased in SIM2i MCF10A cells as compared to Scr controls and is consistent with an EMT-like phenotype (Laffin et al., 2008). Data are represented as mean±s.e.m., asterisk (*) indicates P<0.05; Bar=100 µm.