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. 2015 Feb 23;9:52. doi: 10.3389/fncel.2015.00052

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Copyright © 2015 Tarang, Doi, Gurumurthy, Harms, Quadros and Rocha-Sanchez.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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TetO-driven, Dox-activated expression of the DN-CBRb transgene leads to downregulation of the endogenous RB1 in vitro. HEK293 cells, which endogenously express RB1 (Chano et al., 2006), were co-transfected with TetO-DN-CB-myc₆-Rb1, and the pCMV-Tet3G transactivator vector. In the absence of Dox, co-transfected HEK293 cells stably expressed RB1 (lane 1). Addition of Dox to the system for a 24-h period led to TetO-DN-CB-myc₆-Rb1 activation and RB1 downregulation (lane 2). Culturing a subset of those Dox-treated cells in a Dox-free media for an additional 24 h allowed for RB1 expression to be resumed (lane 3). C = Control HEK293 cells transfected with pCMV-Tet3G only, treated with Dox.