dLKR/SDH interacts directly with the N-terminal region of H3 and H4 and represses CARMER-mediated transcription. (A) 2 μg of recombinant GST or GST-LKR was incubated with calf thymus histones and pulled down using glutathione Sepharose beads, and half of the pull-down was separated by SDS-PAGE and blotted with histone H3 antibody. The other half was stained with Coomassie Brilliant blue (bottom), showing both histone H3/H4, GST, and GST-LKR. The second and fourth lanes do not contain NADH and α-ketoglutarate, and the fifth lane is calf thymus bulk histones alone to show the migration of H3, H2B, H2A, and H4. GST and GST LKR is shown. (B) 10 μg of histones was incubated with 2 μg of GST-LKR in the presence of increasing amounts of histone peptide competitors corresponding to histone H3 amino acids 1–10, 10–20, and 74–83. Pull-downs were performed and blotted for histone H3. One half of the pull-down was stained with Coomassie Brilliant blue showing GST-LKR. (C) Transfections and luciferase assays were performed as in Fig. 2 using 1 μg pEcR-Luc reporter and 0.4 μg pIE-CARMER. Where indicated, ethanol (−) or ecdysone (Ecd +) was used. Error bars represent SD.