Mouse anti phospho histone 3

Coverslips were fixed in 4% PFA and immunostained with rabbit anti-Olig2 (EMD Millipore, Billerica, MA), rat anti-MBP (Biorad), mouse anti-phospho-histone 3 (Cell Signaling, Danvers, MA), or mouse anti-Nk×2.2 (Developmental Hybridoma Bank, University of Iowa). Secondary fluochrome-tagged antibodies were obtained from (Invitrogen/Thermo Fisher, Waltham, MA). Images were obtained on an Axioimager Z1 microscope (Zeiss, Oberkochen, Germany). Concerning ex-vivo experiments, P5 and P30 mouse brains were collected in the 4 experimental groups designed (PBS, Nimesulide, IL-1β, IL-1β+Nimesulide) and fixed to obtain 10μm thick coronal sections. Immunostainings with rabbit anti-NG2 (Millipore) on P5 brains to quantify OPCs and mouse anti-MBP (Millipore) antibodies on P30 brains for myelinated axons were performed as previously described (Favrais et al., 2011 (link)). NG2+ cells were counted within the white matter tracts of the external capsule using ImageJ software (NIH, Bethesda, MD). MBP immunostaining intensity was assessed by ImageJ densitometry analysis at the level of the sensory-motor cortex.