(A) Anti-flag immunoblot of nuclear extract prepared from HeLa cells transiently transfected with: empty flag vector (lane 1), FlagMaf1 (lane 2), FlagBrf1 (lane 3), FlagBrf2 (lane 4)

<p><b>Copyright information:</b></p><p>Taken from "Human Maf1 negatively regulates RNA Polymerase III transcription via the TFIIB family members Brf1 and Brf2"</p><p></p><p>International Journal of Biological Sciences 2007;3(5):292-302.</p><p>Published online 1 May 2007</p><p>PMCID:PMC1865091.</p><p>© Ivyspring International Publisher. This is an open-access article distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by-nc-nd/3.0/). Reproduction is permitted for personal, noncommercial use, provided that the article is in whole, unmodified, and properly cited.</p> Arrows depict location of each protein as determined by anti-Flag antibody. Asterisks denote polypeptide doublet routinely observed in HeLa cells over-expressing FlagMaf1. (B) Same immunoblot from 2A reprobed using anti-actin antibody as loading control for transfections with FlagBrf1, FlagBrf2 and empty Flag vector. (C) Nuclear extract prepared from HeLa cells over-expressing FlagMaf1 treated with calf intestine phosphatase (CIP) for increasing amounts of time and immunoblotted with an anti-Flag antibody. (D) Transient transfection of HeLa cells with pGL3-VAI (100ng) (left panel) or pGL3-U6 (100ng) (right panel) with increasing amounts of FlagMaf1(50ng, 100ng, 200ng) and empty vectors Flag (200ng) and pGL3(100ng) as indicated by + sign