Sample 4
Mus musculus
Protocols: Mice. Colonies of LysM-Bmal1-/- and CX3CR1-Bmal1-/- mice were maintained in the University of Manchester Biological Services Facility (BSF). Where WT only mice were used C57BL/6 were purchased from Envigo. All genetically modified mice were created on a C57BL/6 background and housed in 12:12 light/dark (L:D) cycles with food and water supplied ad libitum. All protocols were approved by the University of Manchester Animal Welfare and Ethical Review Body and the Animals (Scientific Procedures) Act 1986 UK Home Office guidelines were strictly adhered to. Conditional targeted cre positive mice, were sex and age matched with floxed/floxed littermate controls; PECs for ex vivo were harvested from male mice. Genotyping was performed on all experimental animals. Primary Cell Culture. PECS from LysM-Bmal1-/- and littermate controls (n=5) were collected from mice by peritoneal lavage. PECS were isolated resuspended in complete RPMI and seeded at 5x105 cells per well in a 24 well plate. After washing peritoneal macrophages were lysed and RNA extracted with ReliaPrep RNA cell miniprep (Promega). RNA was extracted with ReliaPrep RNA cell miniprep (Promega). Truseq stranded mRNA Assay. Paired end.