Manuscript #2005-04-04108B
Supplementary Figure Legends
Supplementary Figure 1 CoREST enhances histone demethylation. a, Demethylation assay on histone using different combination of recombinant proteins. “1x” corresponds to 200 ng of BHC110, 100 ng of HDAC1, 100 ng of CoREST. b, Demethylation assay on histone using BHC110 (800 ng), HDAC1 (400ng), CoREST (400ng) in the absence or presence of HDAC inhibitors (Na+ butyrate, 100mM; TSA, 100ng/ml).
Supplementary Figure 2 Both SANT domains are involved in nucleosomal demethylation. a, Generation of five bacterial expression constructs for CoREST and its deletions containing N-terminal FLAG tag (DELM2, DSANT2, DSANT1 and DSANT1+2). b, Analysis of bacterial recombinant proteins for CoREST and its deletions by colloidal staining. c, Coimmunoprecipitation analysis between BHC110 (1 mg) and either bacterial recombinant CoREST or its deletions (approximately molar amounts equal to that of BHC110) using anti-BHC110 antibodies. d. Demethylation assay on nucleosome using recombinant BHC110 (800ng) and either bacterial recombinant CoREST or its deletions (approximately molar amounts equal to that of BHC110).
Supplementary Figure 3 BHC110 depletion increases mRNA levels of Synapsin and muscarinic Acetylcholine receptor 4. Levels of mRNA were analyzed by quantitative RT-PCR after treatment of HEK 293 cells with siRNA against BHC110. siRNA against luciferase was used as control.
1