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Figure 3 | Breast Cancer Research

Figure 3

From: A novel approach for the generation of genetically modified mammary epithelial cell cultures yields new insights into TGFβ signaling in the mammary gland

Figure 3

Functional characterization of immortalized mammary epithelial cell cultures. (a) Whole mount of a reconstituted mammary gland generated by transplantation of 2.5 × 104 immortalized mammary epithelial cells (IMECs) into a cleared mammary fat pad. (b) PCR analysis of β-casein mRNA expression as an indicator of lactogenic differentiation. IMECs growing in two-dimensional culture under nonpermissive conditions were exposed to lactogenic hormones for 3 days prior to isolation of mRNA for analysis. RNA from HC11 cells treated in the same way and RNA isolated from a mid-pregnant mammary gland were used as positive controls. (c) Responsiveness to transforming growth factor beta (TGFβ)-mediated growth inhibition in IMECs and primary mammary epithelial cells (MECs) was determined under nonpermissive conditions by assessing incorporation of [3H]thymidine. Results are means of triplicate determinations. IMEC-A and IMEC-B are independently generated IMEC cultures derived from different mice, isolated several months apart. The responses of early (P20) and late (P64) passage cultures of IMEC-A were compared.

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Breast Cancer Research

ISSN: 1465-542X

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