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Fig. 3 | Breast Cancer Research

Fig. 3

From: High AUF1 level in stromal fibroblasts promotes carcinogenesis and chemoresistance and predicts unfavorable prognosis among locally advanced breast cancer patients

Fig. 3

Ectopic expression of AUF1 in BSFs promotes carcinogenesis in a paracrine manner. A TCF-64 cells were transfected with vectors either empty (TCF64-CTL) or bearing p37AUF1-ORF (TCF64-ORF). Whole cell lysates were prepared from these cells and were used for immunoblotting using antibodies against the indicated proteins. B TCF64-CTL and TCF64-ORF cells were cultured in SFM for 24 h, and the respective SFCM (TCF64-CTL-SFCM and TCF64-ORF-SFCM) were collected and were used to treat the indicated BC cells for 24 h. Whole cell lysates were prepared and were used for immunoblotting analysis. The numbers below the bands represent fold change relative to the corresponding control after correction against the internal control, GAPDH. C Exponentially growing cells were added in SFM to the upper wells of the CIM plates either separated by a Matrigel basement membrane matrix (invasion) or without (migration), or E-plate (proliferation), and then, these capacities were assessed for the indicated periods of time using the RTCA-DP xCELLigence System. Data are representative of different experiments performed in triplicate. D Cells were double-stained for CD24 and CD44, and the proportions of CD44high/CD24low subpopulations were determined by flow cytometry and are indicated in the boxes as mean ± SD.; n = 3. E Cells (1000) were cultured in ultra-low attachment 96-well plates in the presence of specific stem cell medium. Left panel, representative images of mammospheres. Scale bar, 50 μm. Right panel, Graphs depicting the number of formed mammospheres. Experiments were performed in triplicate, and error bars represent means ± SD (n = 3) (*P < 0.05)

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