Olig2 antibody

Rat OPCs were isolated from rat pups by immunopanning as previously described (Dugas and Emery, 2013 (link)). The cells were transfected with a plasmid encoding the FLAG-tagged ZFP24. Immunoprecipitation was performed using anti-Flag antibody (Sigma-Aldrich). Immunoprecipitation and isolation of the ZFP24-bound DNA was performed using the DNA ChIP-IT Express kit (Active Motif) according to the manufacturer’s instructions. The immunoprecipitated DNA was amplified using iQ SYBR Green Supermix (Bio-Rad) in a Bio-Rad C1000 thermocycler. OLIG2 is known to bind upstream to p21 (Ligon et al., 2007 (link)). We used this known protein-DNA interaction as a control. OLIG2 was immunoprecipitated using OLIG2 antibody (R&D). The following set of primers were used: Sox10: distal: forward: 5′-GCAGAGGACATGGTTGTGGA-3′, reverse: 5′-CTGCCTCAGACTGCTGACAA-3′. Sox10: no TCAT: forward: 5′-CA TACCCAAGGGCTCTCTGC-3′, reverse: 5′-TCCACAACCATGTCCTCTGC-3′. Sox10: TCAT: forward: 5′-GGGTAGGTCACAACCCACTG-3′, reverse: 5′-TG CCATCCTTTGTGTGTGGT-3′. Zfp536: forward: 5′-CGGCTCCTCTAACGT GACTG-3′, reverse: 5′-CCTCGCCGATGTCTGAAGAA-3′. Elovl7: forward: 5′-TGCTTATACAGCGTGCACCA-3′, reverse: 5′-TGGACTGAGGATTGAACT CAGA-3′. Smad7: forward: 5′-AGGATCTTGTCCCCGAGCAG-3′, reverse: 5′-CTGCGTCTCAGGCAGCTCTC-3′. p21: forward: 5′-AGGTGTCTAGACTC CAGATT-3′, reverse: 5′-AAAATCAAGGCTTTGCTGG-3′.