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Fig. 3 | Breast Cancer Research

Fig. 3

From: Role of ADAM17 in the non-cell autonomous effects of oncogene-induced senescence

Fig. 3

Contribution of ADAM17 to the p95HER2-induced senescence secretome. a MCF7 Tet-Off p95HER2 cells constitutively expressing a control shRNA (NT, non-targeting) or two independent shRNAs targeting ADAM17 (A17#1 and A17#2) were cultured with or without doxycycline for 7 days. Cells were then harvested and lysed, and cell lysates were analyzed by Western blotting with the indicated antibodies. b Senescence-associated β-galactosidase was analyzed in the same cells as in (a). Representative bright-field images are shown. c The same cells as in (a) were plated with or without doxycycline and counted at the indicated time points. The results represent averages of two independent determinations. d The conditioned media of the same cells as in (a), treated without doxycycline, were analyzed by label-free quantitative proteomics in triplicate (a-c). The heatmap shows log2FC of the normalized levels of soluble ectodomains. e-g The same cells as in (a) were treated with or without doxycycline for 7 days. Conditioned media from the last two days and cell lysates were analyzed by enzyme-linked immunosorbent assay (e) or Western blotting (f, g) as indicated. Quantification of at least three independent experiments is represented as average and standard deviation. These results were reproduced by using the second stable cell line expressing an independent shRNA targeting ADAM17 (data not shown). Doxy doxycycline

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