Osteoblastic loss of Ezh2

Osteblastic loss of Ezh2 was achieved by breeding Ezh2flox/flox (Su et al., PMID: 12496962) with Osx-Cre (Rodda et al., PMID:16854976). Overall design: Calvaria were dissected out from 2 to 3 day old wild type (WT, Ezh2_WT_Osx) and Ezh2 conditional knockout (Ezh2 cKO, Ezh2_cKO-Osx), washed in PBS, digested for 20 minutes in collagenase digestion medium (aMEM, 2mg/ml Collagenase type II, and 0.005% trypsin) at 37oC, washed in PBS, and snap frozen with liquid nitrogen and stored at -80oC. RNA was then isolated utilizing Ultra Turrax T25 (IKA) and Direct-zol™ RNA kit (Zymo Research).