Fig. 2

Prevention of microglial activation during dissociation and cell sorting protocols using transcription and translation inhibitors. A Brain tissue from adult mice was dissociated using Miltenyi’s ABDK protocol and CD11b-positive cells were enriched using MACS CD11b magnetic microbeads. Purity of CD11b-positive population was controlled on a pool of cells from each sample by cytometry. B, C IL-1β (B) and TNFα (C) mRNAs were quantified using calibrated RT-qPCR following tissue dissociation and CD11b MACS cell enrichment from adult mouse brain (n = 2). Dissociation and MACS protocols were performed with classic buffer (ø), with buffer complemented with transcription inhibitor ActD (3 µM) or with buffer complemented with both transcription and translation inhibitors ActD, Anis and Trip (3 µM, 100 µM and 10 µM, respectively). Transcript levels are expressed as fold change of the classic buffer (ø) condition. Individual values (dots) and average values (bars) are presented