1. MED12 interacts with TDRD3 in a CARM1-dependent fashion.
MED12 interacts with TDRD3 in a CARM1-dependent fashion. (A) Fluorograph (top panel) and Coomassie Brilliant Blue staining (bottom panel) of the peptides in vitro methylated by recombinant CARM1 in the presence of tritium-labeled AdoMet. (B) The peptides were used to pull down Tudor domains of the indicated proteins. The input samples and the eluted samples were immunoblotted with αGST antibody. Streptavidin HRP serves as a peptide loading control. (C) MCF-7-Tet-on-shCARM1 cells were untreated or treated with doxycycline (1 μg/ml) for 8 d. Nuclear extracts were subjected to IP with αTDRD3 antibody and the eluted samples were detected by Western blotting with αMED12 and αTDRD3. The input samples were immunoblotted with αTDRD3, αMED12, αmeMED12a, and αCARM1. (D) HEK293T cells were transiently transfected with FLAG, FLAG-MED12 WT, FLAG-MED12-R1862K, FLAG-MED12-R1899K, and FLAG-MED12-R1862,1899,1912K. Total cell lysates were immunoprecipitated with αTDRD3 antibody and the eluted samples were subjected to Western analysis with αFLAG and αTDRD3 antibodies. The input samples were immunoblotted with αFLAG, αTDRD3, and αβ-actin.
Donghang Cheng et al. LSA 2018;1:e
© 2018 Cheng et al.