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. 2021 Feb 5;13:1123–1133. doi: 10.2147/CMAR.S264414

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© 2021 Savukaitytė et al.

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Different effects of REDD1 silencing on aspirin-mediated expression of phosphorylated 4E-BP1. Cells were transfected with non-targeting (siCT) or REDD1 siRNA (siREDD1) for 24 (MDA-MB-468, MCF-7, MDA-MB-231 cell lines) or 48 hours (MCF10A) and treated with vehicle control (C) or 2 mM of aspirin (ASA) for the next 24 hours before cell lysis. Equal amounts of proteins (50 µg) were loaded on each lane of SDS-PAGE gel for electrophoresis, followed by transfer onto PVDF membranes, which were then probed with respective antibody. β-actin was used as loading control. REDD1 was probed on a different blot than p-4E-BP1. Data are expressed as means±SD from three independent experiments for MDA-MB-468, MCF-7, MDA-MB-231, and from two independent experiments for MCF10A.