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Fig. 8. MASH1 binds to the I12b enhancer in vivo. LGE and MGE were
dissected from E11.5 mouse telencephalon, cross-linked and immunoprecipitated
using an anti-MASH1 antibody (
-Mash1). PCR analysis was performed on
the immunoprecipitated chromatin using primers corresponding to the I12b
enhancer (I12b Enh) and the Hes6 upstream enhancer (Hes6 Enh). After
28-30 cycles, PCR products (605 bp for mI12b and 176 bp for Hes6) were
electrophoresed on 1% agarose gel. As controls, amplifications were carried
out on chromatin obtained from the elution of protein-A sepharose beads that
was preincubated with the cross-linked chromatin or with the IP dilution
buffer (No antibody and Mock, respectively). Different dilutions of total
input chromatin (Genomic DNA) were used for each set of primers: 1/500 for
I12b and 1/1000 for Hes6 Enh.