Prdm12 binds dbx1 and nkx6 promoter regions, promotes their methylation and affects the activity of a dbx1 promoter-lacZ reporter construct. (A) IGV browser screenshots showing differential binding of Flag-mPrdm12 in RA-treated neuralized caps versus naïve caps in the promoters of dbx1, nxk6-1 and nkx6-2. Two major peaks in the dbx1 locus (at −3 kb and −5 kb), one in the nkx6-1 locus (at −0.5 kb), and another in the nkx6-2 locus (at −1.0 kb) are boxed. (B) ChIP-qPCR analysis of the dbx1, nkx6-1 and nkx6-2 promoter regions (with primers designed at the dashed boxes highlighted in A) in RA-treated neuralized caps overexpressing or not mPrdm12 with α-H3K9me2 (upper histogram) or α-H3K9me3 (lower histogram) antibodies. Fold enrichment is shown. (C) X-gal staining of a transverse section of the neural tube of a chick embryo electroporated with a 5.7 kb mouse eDbx1::lacZ reporter, plus or minus a pCIG-Prdm12 or an empty pCIG vector, and immunostained for GFP to show electroporation efficiency. Quantification of the number of X-gal+ cells observed in the electroporated side is shown (n≥4 embryos). Error bars denote s.e.m. **P<0.01.