Illumina HiSeq 1500 sequencing; ChIP-seq of Pbx1, Prep1 and Sp2 in WT MEFs, Sp2ko MEFs and in Sp2ko MEFs ectopically expressing Flag-tagged full length Sp2; and ChIPexo of Sp2 in WT and  Sp2ko MEFs
ChIP-seq of Pbx1, Prep1 and Sp2 in WT MEFs, Sp2ko MEFs and in Sp2ko MEFs ectopically expressing Flag-tagged full length Sp2; and ChIPexo of Sp2 in WT and  Sp2ko MEFs
cell culture, see growth and nucleic acid extraction protocol. Antibodies used: Anti-Pbx1 (Cell Signaling Technology, Danvers, USA, Cat. No. #4342); Anti-Prep1 (Santa Cruz, Dallas, USA, Cat. No. sc-25282); antiSp2 Ab1 (produced in-house) MEFs were cultured in a 1:1 mixture of Dulbecco's modified eagle medium (DMEM)/high glucose (4.5 g/L) (Gibco Thermo Fisher Scientific, Waltham, USA) and HAM's F-10 (Gibco Thermo Fisher Scientific, Waltham, USA) supplemented with 10% (v/v) fetal calf serum (Sigma-Aldrich, St. Louis, USA) and 1% Penicillin-Streptomycin. Sp2ko MEFs expressing Sp2 mutants were selected and propagated in the presence of 2 µg/mL puromycin. Conventional ChIPs of Pbx1, Prep1 and Flag-Sp2 were performed using the One Day ChIP kit (Diagenode, Seraing, Belgium). Three microgram of antibody were used per ChIP experiment, and five nanogram of precipitated DNA were used for indexed sequencing library preparation using the Microplex library preparation kit v2 (Diagenode). Microplex library preparation kit
Experimental Factor: immunoprecipitate
anti-Pbx1
Experimental Factor: genotype
Sp2 homozygous knockout