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. 2012 Jun 25;32(18):2261–2272. doi: 10.1038/onc.2012.252

Figure 3.

Figure 3

Beclin 1 expression is upregulated in mammospheres.(a) The single-cell suspension from either fresh breast cancer samples or various different breast cancer cell lines was cultured under adherent or mammosphere conditions. The expression of the BECN1 gene in mammospheres and adherent cells was investigated using RT–qPCR. Results are expressed as the mean±s.d. for three independent determinations and analyzed using Student's t-test (*P<0.05, **P<0.01, ***P<0.001). The fold change was calculated relative to control adherent cells. (b) Primary adherent cells and primary mammospheres were incubated in complete medium (control) or with EBSS for 4 h, and then the expression level of the BECN1 gene was determined as in (a) using quantitative RT–qPCR (left). Results are expressed as described in (a). Immunoblot analysis of Beclin 1 levels in primary mammospheres and adherent cells (right). Control was set as in 1 in untreated cells. (c) MCF-7 mammospheres and adherent cells were incubated in complete medium or with EBSS for the times indicated, and then the expression of BECN1 gene was determined as in (a, left). Results are expressed as described in (a). Immunoblot analysis of Beclin 1 levels in MCF-7 mammospheres and adherent cells (right).