Expression data from vehicle tumor associated macrophages (TAM) VS PLX3397 treated-TAM.. Mus musculus

To test the difference genes expression of TAMs obtained from orthotopically implanted hepa1-6 tumors in C57BL/6 mice.Isolated TAMs were obtained from orthotopically implanted hepa1-6 tumors in C57BL/6 mice. Specimens were minced with scissors and digested by incubation for 1 h at 37°C in high-glucose DMEM (Life Technologies, CA) containing 0.1% collagenase IV (Sigma). After being washed in medium plus 10% FBS, the cell suspension was forced through a graded series of meshes to obtain single-cell suspensions and centrifuged at 400 g for 30–40 minutes at 18–20°C on layers of Lympholyte-M (Cedarlane Labs, Burlington, Ontario, Canada), after which the upper layer was removed, thus leaving the lymphocyte layer undisturbed at the interface. TAMs were sorted using Fluorescence-activated cell sorting (FACS, Becton Dickinson, Franklin Lakes, NJ) with CD45+, CD11b+, and F4/80+ (Becton Dickinson) according to the protocol listed at http://www.immgen.org/Protocols/ImmGen%20Cell%20prep%20and%20sorting%20SOP.pdf . TAMs growing in DMEM supplemented with 10% FBS were tested to determine their biological characteristics. We used microarrays to detail the global programme of gene expression and identified vehicle-TAM and treated-TAM during this process. Overall design: To test the difference genes expression of vehicle-TAM and PLX3397 treated-TAM.TAMs from mouse tumors were processed in Trizol within 4 h after sorting and then analyzed using whole-mouse genome Affymetrix Mouse Gene 1.0 ST Arrays.