Anti rarγ

ChIP was performed according to the manufacturer’s protocol (Active Motif) as described (Frank et al., 2001 (link)). To perform whole-embryo ChIP, nuclear extracts were prepared from pooled wild-type E8.25 mouse embryos as previously described (Kumar and Duester, 2014 (link)); antibodies included anti-RARα or anti-RARγ (Santa Cruz Biotechnology) or anti-RARβ (Affinity Bioreagents). ChIP samples were subjected to PCR using primers flanking the mouse Wnt8a RARE near −4.9 kb (5′-ATC TTG GGT TGA GGC AGA GTC TC-3′ and 5′-CGC TGA GCC ACC TCT ACA ATC TT-3′ that generate a 280bp PCR product) or primers flanking a non-specific region located at −9.7 kb (5′-TCA GAC ATG GCC TCC ACT AGA AC-3′ and 5′-CCC CGT TGA GGT ATT TCT TTT GGC-3′ that generate a 219bp PCR product).