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. 2016 Mar 24;6:23727. doi: 10.1038/srep23727

Figure 1. The role of DynII in ERα nuclear and extra-nuclear activities and in E2-induced cell proliferation.

Figure 1

(a) Western blotting and relative densitometric analyses (a’, a”) of AKT and ERα S118 phosphorylation in MCF-7 control (CTR) and DynII knock-down cells treated with E2 (10 nM) at different time points. (b) ERα:IGF-1R co-immunoprecipitation and relative densitometric analyses (b’) in MCF-7 control (CTR) and DynII knock-down cells treated with E2 (10 nM) at the indicated time points. The loading control was done by evaluating vinculin expression in the same filter. *indicates significant differences with respect to the control (0) sample; °indicates significant differences with respect to the corresponding E2 sample. (c) RT-qPCR analysis of pS2/TIFF (pS2), progesterone receptor (PR) and cathepsin D (Cat D) mRNA expression normalized to the GAPDH mRNA expression in MCF-7 control (CTR) and DynII knock-down cells treated with E2 (10 nM) for 24 hrs. *indicates significant differences with respect to the control (CTR−); °indicates significant differences with respect to the E2 CTR sample. Western blotting (d) and relative densitometric (d’) analyses of pS2/TIFF (pS2), progesterone receptor (PR), cathepsin D (Cat D), cyclin D1 (Cyc D1) and Bcl-2 expression levels in MCF-7 control (CTR) and DynII knock-down cells treated with E2 (10 nM–24 hrs). The loading control was done by evaluating tubulin or vinculin expression in the same filter. *indicates significant differences with respect to the control (CTR−) sample; °indicates significant differences with respect to the corresponding E2 CTR sample. (e) The number of MCF-7 control (CTR) and DynII knock-down cells treated with E2 (10 nM–24 hrs). *indicates significant differences with respect to the control (−); °indicates significant differences with respect to the E2 sample; Time 0 corresponds to plated cells. (f) Representative distribution of two experiments performed in MCF-7 control (CTR) and DynII knock-down cells in the different phases of the cell cycle and relative quantitation (f’).