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

Figure 3. The role of autophagic flux on neo-synthesized and mature ERα.

Figure 3

(a) Western blotting analysis and relative densitometric analyses (a’) of ERα cellular levels in MCF-7 cells treated with bafilomycin A1 (Baf) (100 nM–2 hrs) evaluated in the presence or absence of cycloheximide (CHX) (1 μg/ml–6 hrs). (b,c) Western blotting analysis and relative densitometric analyses of mature ERα content in MCF-7 cells kept in methionine-free medium for 24 hrs and treated for 2 hrs with E2 (10 nM) in the presence or in the absence of bafilomycin A1 (Baf) (100 nM–2 hrs) with or without cycloheximide (CHX) (1 μg/ml–6 hrs) administration. The loading control was done by evaluating vinculin expression in the same filter. *indicates significant differences with respect to the control (−) sample; °indicates significant differences with respect to the corresponding E2 sample. (d) Immunoprecipitation analysis of neo-synthesized ERα cellular levels in MCF-7 cells treated for 2 hrs with E2 (10 nM) in the presence or absence of bafilomycin A1 (Baf) (100 nM–2 hrs). (d’) Western blotting analysis of biotin-labelled cellular proteins. The loading control was done by evaluating vinculin expression in the same filter.