Fig. 4.
TAL1 recruits LSD1 to the p4.2 promoter in undifferentiated, but not differentiated, MEL cells. (A) Schematic representation of the mouse P4.2 gene locus. The primers are indicated. (B) mRNAs from WT or TAL1-transduced MEL cells treated with 1.5% DMSO were purified and analyzed by Northern blotting using the P4.2 probe. (C and D) Cross-linked chromatin from MEL cells in the presence and absence of 1.5% DMSO was precipitated with TAL1 antibody. The TAL1-chromatin complexes were reversed and the selected DNA was analyzed by PCR (C). (D) TAL1-DNA complexes from the first TAL1 ChIP were subjected to a second IP with anti-LSD1. The cross-links of these double IP protein/DNA complexes were reversed, and the DNA was analyzed by PCR. (E) Cross-linked chromatin from MEL cells in the presence and absence of 1.5% DMSO was precipitated with TAL1 and LSD1 antibodies. The selected chromatin complexes were reversed and analyzed by quantitative PCR. (F) The cross-linked chromatin from MEL cells treated with or without DMSO was precipitated with dimeH3K4 antibody. The precipitated DNA fragments were amplified by using primers specific for −6 Kb, the P4.2 promoter, and 3′ UTR regions by quantitative PCR.