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Figure 2 | Breast Cancer Research

Figure 2

From: Type I insulin-like growth factor receptor over-expression induces proliferation and anti-apoptotic signaling in a three-dimensional culture model of breast epithelial cells

Figure 2

Lack of acinar overgrowth in MCF-10A cells expressing kinase-dead IGFIR. (a) Differential interference contrast images of acinar structures formed from stable pools of MCF-10A cells infected with virus expressing empty vector (pCLXSN; panels a, b), kinase-dead (K1003A; panels d, e) and wild-type IGFIR (panels g, h); scale bar = 80 μm. Equatorial confocal sections of pCLXSN (panel c), K1003A (panel f) and wild-type IGFIR (panel i) acinar structures labeled with antibodies to the IGFIR (green) and the nuclear counterstain TO-PRO-3 (blue); scale bar = 40 μm. Cells were grown for 20 days in media supplemented with 10 μg/ml insulin (panels a, c, d, f, g, i) or 100 ng/ml IGF-I (panels b, e, h). (b) Western blots of whole-cell lysates from monolayer cultures of MCF-10A cells expressing empty vector (pCLXSN), kinase-dead (K1003A) or wild-type IGFIR (IGFIR) untreated or treated with 50 ng/ml IGF-I (left panel) or 10 μg/ml insulin (right panel) for 5 minutes, 60 minutes or overnight (o/n) after overnight incubation in serum-free media. Proteins were separated by 12.5% SDS-PAGE, transferred to PVDF and incubated overnight with the indicated antibodies to signaling proteins downstream of IGFIR activation.

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Breast Cancer Research

ISSN: 1465-542X

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